http://2013.igem.org/wiki/index.php?title=Special:Contributions/Surine&feed=atom&limit=50&target=Surine&year=&month=2013.igem.org - User contributions [en]2024-03-28T18:28:16ZFrom 2013.igem.orgMediaWiki 1.16.5http://2013.igem.org/Team:USTC_CHINA/TeamTeam:USTC CHINA/Team2013-09-27T23:35:45Z<p>Surine: </p>
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<h2>Students</h2><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/2/21/Zhangsitao1.JPG" alt="Zhang Sitao"> <br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/0/0d/Zhangsitao2.JPG" alt="Zhang Sitao"><br />
<div class ="nameplate"><a href="#">Zhang Sitao</a> </div><br />
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<img src="https://static.igem.org/mediawiki/2013/0/0d/Zhangsitao2.JPG" alt="Zhang Sitao" align="left"><p>Our labor leader weighs various matters, leads the overall trend and plays our cards right. He leaves a strong impression in others' mind. However, His friends found that the leader is very cute.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/0/02/Zhaochanglong1.JPG" alt="Changlong Zhao"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/7/7e/Zhaochanglong2.JPG" alt="Changlong Zhao" ><br />
<div class ="nameplate"><a href="#">Changlong Zhao</a></div><br />
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<img src="https://static.igem.org/mediawiki/2013/7/7e/Zhaochanglong2.JPG" alt="Changlong Zhao" align="left"><p>There's no doubt that we can give full stars for Changlong's fighting capacity. The roads leading to success will never be smooth and Changlong is a perfect companion to travel with.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/9/91/Xionghanjin1.JPG" alt="Hanjin Xiong"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/d/d9/Xionghanjin2.JPG" alt="Hanjin Xiong" ><br />
<div class ="nameplate"><a href="#">Hanjin Xiong</a></div><br />
<div class = "details"><br />
<img src="https://static.igem.org/mediawiki/2013/d/d9/Xionghanjin2.JPG" alt="Hanjin Xiong" align="left"><p>As the keynote speaker of our team, he always keeps a clear head with extraordinary creativity and expressiveness. He said, "It is shameful if you haven't burnt the midnight oil for iGEM." Moving forward bravely, he shows us overwhelming power which nobody can stop it.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/8/85/Shenshengqi1.JPG" alt="Shen Shengqi"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/4/44/Shenshengqi2.JPG" alt="Shen Shengqi" ><br />
<div class ="nameplate"><a href="#">Shen Shengqi</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/4/44/Shenshengqi2.JPG" alt="Shen Shengqi" align="left"><p>Everyone considers that it is honored to be a friend of "Brother Face" as he is a totally "local tyrant". Actually," Brother Shen" is warmth, nice, really expert in digging shortcuts in the experiments. He is a sharp soldier of our team as he adheres to the "more with less" principle.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/6/60/Shaoxueying1.JPG" alt="Shao Xueying"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/e/e9/Shaoxueying2.JPG" alt="Shao Xueying"><br />
<div class ="nameplate"><a href="#">Shao Xueying</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/e/e9/Shaoxueying2.JPG" alt="Shao Xueying" align="left"><p>Competent and independent, Shao Xueying has unique ideas about colors and graphics. At the same time, she is a good lecturer. She edits our wiki and does presentation for us. </p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/f/f9/Yuanye1.JPG" alt="Yvette Yuan"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/8/81/Yuanye2.JPG" alt="Yvette Yuan" ><br />
<div class ="nameplate"><a href="#">Yvette Yuan</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/8/81/Yuanye2.JPG" alt="Yvette Yuan" align="left"><p>As a s pronoun for efficient, Yuan Ye is studious and decisive. And what makes her best is that she always brings us delicious oranges.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/f/f3/Yanggege1.JPG" alt="Gege Yang"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/2/25/Yanggege2.JPG" alt="Gege Yang"><br />
<div class ="nameplate"><a href="#">Gege Yang</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/2/25/Yanggege2.JPG" alt="Gege Yang" align="left"><p>She is a lovely girl that everyone likes her, she is also a good experimenter that every step of our project has her contribution.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/2/23/Limingyue1.JPG" alt="Mingyue Li"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/0/05/Limingyue2.JPG" alt="Mingyue Li"><br />
<div class ="nameplate"><a href="#">Mingyue Li</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/0/05/Limingyue2.JPG" alt="Mingyue Li" align="left"><p>"Mingyue Bacteria" is the spokesperson of our bacterium, handling the destiny of those little lives. We all agreed that, Mingyue with rubber gloves is GORGEOUSNESS!</p></div><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/c/c1/Madanyi1.JPG" alt="Danyi Ma"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/1/19/Madanyi2.JPG" alt="Danyi Ma" ><br />
<div class ="nameplate"><a href="#">Danyi Ma</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/1/19/Madanyi2.JPG" alt="Danyi Ma" align="left"><p>"Aunty Ma" takes charge of our finance and safety, which calls for much patience and responsibility. In the experiment, she also plays an absolutely necessary role.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/7/76/Panminghao1.JPG" alt="Minghao Pan"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/e/ed/Panminghao2.JPG" alt="Minghao Pan" ><br />
<div class ="nameplate"><a href="#">Minghao Pan</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/e/ed/Panminghao2.JPG" alt="Minghao Pan" align="left"><p>Modeling, modeling, and modeling. PS: Taking photos is really not my cup of tea.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/8/87/Zhangheng1.JPG" alt="Zhang Heng"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/6/63/Zhangheng2.JPG" alt="Zhang Heng" ><br />
<div class ="nameplate"><a href="#">Zhang Heng</a> </div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/6/63/Zhangheng2.JPG" alt="Zhang Heng" align="left"><p>He is a man full of responsibility, we could 100% trust him! Bro, it's you that bring us positive energy!</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/1/18/Chenzhuo1.JPG" alt="Chen Zhuo"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/a/af/Chenzhuo2.JPG" alt="Chen Zhuo"><br />
<div class ="nameplate"><a href="#">Chen Zhuo</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/a/af/Chenzhuo2.JPG" alt="Chen Zhuo" align="left"><p>Always, he is still of tongue, but he is not only a genius of experiment, but also a brilliant living library. I cannot say more but admire! </p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/3/3f/Chenzhaoxiong1.JPG" alt="Zhaoxiong Chen"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/d/dc/Chenzhaoxiong2.JPG" alt="Zhaoxiong Chen"><br />
<div class ="nameplate"><a href="#">Zhaoxiong Chen</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/d/dc/Chenzhaoxiong2.JPG" alt="Zhaoxiong Chen" align="left"><p>This smart boy is good at playing all kinds of computer systems. We believe that he will refresh the history of wikis!</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/1/15/Wuming1.JPG" alt="Min Wu"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/b/be/Wuming2.JPG" alt="Min Wu"><br />
<div class ="nameplate"><a href="#">Min Wu</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/b/be/Wuming2.JPG" alt="Min Wu" align="left"><p>I love experiments. I love games. I love Weibo.</p><br />
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<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/d/df/Pengyali1.JPG" alt="Yali Peng"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/0/0c/Pengyali2.JPG" alt="Yali Peng"><br />
<div class ="nameplate"><a href="#">Yali Peng</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/0/0c/Pengyali2.JPG" alt="Yali Peng" align="left"><p>She is a quiet girl, she likes smile, she loves doing experiments peacefully and slowly. As the best partner of Mingyue Li, every trouble become easy! Hey, little pretty we all love you! </p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/4/4a/Xuehao1.JPG" alt="Hao Xue"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/0/08/Xuehao2.JPG" alt="Hao Xue"><br />
<div class ="nameplate"><a href="#">Hao Xue</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/0/08/Xuehao2.JPG" alt="Hao Xue" align="left"><p>Laughing, he is still laughing! What on hill? Oh god, negative results, but how… But bro, thank you for giving us positive energy, you really raise us up! My bro! </p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/4/49/Qiuyanning1.JPG" alt="Yanning Qiu"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/d/df/Qiuyanning2.JPG" alt="Yanning Qiu"><br />
<div class ="nameplate"><a href="#">Yanning Qiu</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/d/df/Qiuyanning2.JPG" alt="Yanning Qiu" align="left"><p>Our little sister holds the trump cards. She always knows what do with all the words and pictures. Brave and creative, Yanning enjoys the days with new skills and knowledge. Our team was painted colorfully with the lively girl.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/5/52/Longjie1.JPG" alt="Long jie"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/9/96/Longjie2.JPG" alt="Long jie"><br />
<div class ="nameplate"><a href="#">Long jie</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/9/96/Longjie2.JPG" alt="Long jie" align="left"><p>Clever and hard-working, I cannot agree more to do experiments with him. You never let us down!</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/c/c8/Dongbo1.JPG" alt="Bo Dong"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/0/08/Dongbo2.JPG" alt="Bo Dong" ><br />
<div class ="nameplate"><a href="#">Bo Dong</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/0/08/Dongbo2.JPG" alt="Bo Dong" align="left"><p>He is an earnest boy, he always work hard that every bros and sis like him, he is our team's MVP! <br />
Hey, bro! It our pleasure to be with you!</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/1/14/Wangshiwei1.JPG" alt="Shiwei Wang"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/e/e2/Wangshiwei2.JPG" alt="Shiwei Wang" ><br />
<div class ="nameplate"><a href="#">Shiwei Wang</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/e/e2/Wangshiwei2.JPG" alt="Shiwei Wang" align="left"><p>Again, a quiet boy is coming! He love experiment, he is Bo Dong's loyal friend. We all believe in him, without his help we cannot achieve our goal! Thanks a lot ,my bro! </p><br />
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<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/a/a3/Xinghuayue1.JPG" alt="Huayue Xing"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/a/ae/Xinghuayue2.JPG" alt="Huayue Xing" ><br />
<div class ="nameplate"><a href="#">Huayue Xing</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/a/ae/Xinghuayue2.JPG" alt="Huayue Xing" align="left"><p>With my little eyes, I see bacterium; with my little eyes, I see TD-1; with my little eyes, I see vaccine secreted out; with my little eyes, I see the future without needles. Carefulness, earnest, and a little bit of acting cute, I am XHY.</p><br />
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<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/a/ad/Xionglei1.JPG" alt="Lei Xiong"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/b/b2/Xionglei2.JPG" alt="Lei Xiong" ><br />
<div class ="nameplate"><a href="#">Lei Xiong</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/b/b2/Xionglei2.JPG" alt="Lei Xiong" align="left"><p>Despite the fact that it is me who always breaks test tubes, loses beakers, and pours reagent onto the skin of my hands, I have the enthusiasm for science. I love to explore and pursue knowledge. As long as there is a chance to see the tip of the iceberg, it doesn't matter how many test tubes I am going to break.</p><br />
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<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/b/bd/Caoqinjingwen1.JPG" alt="Cao Qinjingwen"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/f/f9/Caoqinjingwen2.JPG" alt="Cao Qinjingwen"><br />
<div class ="nameplate"><a href="#">Cao Qinjingwen</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/f/f9/Caoqinjingwen2.JPG" alt="Cao Qinjingwen" align="left"><p>Excellent! Without these kinds of words, how can I say anything to describe her? As our elder sister, she always gives us self-confident, hey soul sister!</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/4/4a/Xiaozhuyun1.JPG" alt="Xiao Zhuyun"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/1/1d/Xiaozhuyun2.JPG" alt="Xiao Zhuyun"><br />
<div class ="nameplate"><a href="#">Xiao Zhuyun</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/1/1d/Xiaozhuyun2.JPG" alt="Xiao Zhuyun" align="left"><p>Sincere and straightforward, "piggy", the curve wrecker in our eyes, puts all her efforts into research and study. Only when you get close to her, will you find that she also loves to play, that she also loves life.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/9/9a/Wangzeyu1.JPG" alt="Zeyu Wang"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/5/53/Wangzeyu2.JPG" alt="Zeyu Wang"><br />
<div class ="nameplate"><a href="#">Zeyu Wang</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/5/53/Wangzeyu2.JPG" alt="Zeyu Wang" align="left"><p>I love modeling, too. I hate posing for photographs, neither.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/6/62/Hanyingying1.JPG" alt="Han Yingying"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/c/ca/Hanyingying2.JPG" alt="Han Yingying"><br />
<div class ="nameplate"><a href="#">Han Yingying</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/c/ca/Hanyingying2.JPG" alt="Han Yingying" align="left"><p>Tender as a new-born kitty, Yingying doesn't like to stand in the spotlight. She's a diligent brain instead of a silken tongue. We believe that gold will shine no matter where it is. </p><br />
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</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/e/e4/Fansijia1.JPG" alt="Sijia Fan"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/5/57/Fansijia2" alt="Sijia Fan"><br />
<div class ="nameplate"><a href="#">Sijia Fan</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/5/57/Fansijia2" alt="Sijia Fan" align="left"><p>Black humorist, and sadly, the leader is always shouting at me:" Hurry! Hurry!".</p><br />
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<h2>Advisers</h2> <br />
<div class ="profilewrap" style="width:160px;margin:60px 80px 5px;"><br />
<img style="width:160px" src="https://static.igem.org/mediawiki/2013/f/f0/Haiyan_Liu.jpg" alt="Haiyan Liu"><br />
<div class ="nameplate"><a href="#">Haiyan Liu</a></div><br />
<div class = "details"><img style="width:160px" src="https://static.igem.org/mediawiki/2013/f/f0/Haiyan_Liu.jpg" alt="Haiyan Liu" align="left"><p>Haiyan Liu was born in Sichuan Province, China. He received his BS degree in Biology in 1990 and PhD degree in Biochemistry and Molecular Biology in 1996, both from USTC. Between 1993 and 1995 he was a visiting graduate student in Laboratory of Physical Chemistry of ETH, Zurich (Switzerland). Since 2001, he has been a professor of computational biology at School of Life Sciences, USTC. </p><br />
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<div class ="profilewrap" style="width:160px;margin:60px 80px 5px;"><br />
<img style="width:160px" src="https://static.igem.org/mediawiki/2013/0/09/Hongjiong.PNG" alt="Jiong Hong"><br />
<div class ="nameplate"><a href="#">Jiong Hong</a></div><br />
<div class = "details"><img style="width:160px" src="https://static.igem.org/mediawiki/2013/0/09/Hongjiong.PNG" alt="Jiong Hong" align="left"><p> I am applying this strategy on the mechanism of the complex diseases such as cancer and diabetes. My ongoing project is to identify biomarkers in order to detect the progress stages of the diabetes. In addition, I have planed to analyze the genetic and environmental factors and their interactions during the progressing of the type 2 diabetes with systems-biology approaches.</p><br />
</div><br />
</div><br />
<div class ="profilewrap" style="width:160px;margin:60px 80px 5px;"><br />
<img style="width:160px" src="https://static.igem.org/mediawiki/2013/4/46/Wujiarui.PNG" alt="Jiarui Wu"><br />
<div class ="nameplate"><a href="#">Jiarui Wu</a></div><br />
<div class = "details"><img style="width:160px" src="https://static.igem.org/mediawiki/2013/4/46/Wujiarui.PNG" alt="Jiarui Wu" align="left"><p>Since the research strategy of systems biology is well fit to analyze the biological complex systems. I am applying this strategy on the mechanism of the complex diseases such as cancer and diabetes. We have developed systematic approaches based on proteomics and bioinformatics to analyze human normal and diabetic serum.</p><br />
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</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/TeamTeam:USTC CHINA/Team2013-09-27T23:33:54Z<p>Surine: </p>
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<h2>Students</h2><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/2/21/Zhangsitao1.JPG" alt="Zhang Sitao"> <br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/0/0d/Zhangsitao2.JPG" alt="Zhang Sitao"><br />
<div class ="nameplate"><a href="#">Zhang Sitao</a> </div><br />
<div class = "details"><br />
<img src="https://static.igem.org/mediawiki/2013/0/0d/Zhangsitao2.JPG" alt="Zhang Sitao" align="left"><p>Our labor leader weighs various matters, leads the overall trend and plays our cards right. He leaves a strong impression in others' mind. However, His friends found that the leader is very cute.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/0/02/Zhaochanglong1.JPG" alt="Changlong Zhao"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/7/7e/Zhaochanglong2.JPG" alt="Changlong Zhao" ><br />
<div class ="nameplate"><a href="#">Changlong Zhao</a></div><br />
<div class = "details"><br />
<img src="https://static.igem.org/mediawiki/2013/7/7e/Zhaochanglong2.JPG" alt="Changlong Zhao" align="left"><p>There's no doubt that we can give full stars for Changlong's fighting capacity. The roads leading to success will never be smooth and Changlong is a perfect companion to travel with.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/9/91/Xionghanjin1.JPG" alt="Hanjin Xiong"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/d/d9/Xionghanjin2.JPG" alt="Hanjin Xiong" ><br />
<div class ="nameplate"><a href="#">Hanjin Xiong</a></div><br />
<div class = "details"><br />
<img src="https://static.igem.org/mediawiki/2013/d/d9/Xionghanjin2.JPG" alt="Hanjin Xiong" align="left"><p>As the keynote speaker of our team, he always keeps a clear head with extraordinary creativity and expressiveness. He said, "It is shameful if you haven't burnt the midnight oil for iGEM." Moving forward bravely, he shows us overwhelming power which nobody can stop it.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/8/85/Shenshengqi1.JPG" alt="Shen Shengqi"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/4/44/Shenshengqi2.JPG" alt="Shen Shengqi" ><br />
<div class ="nameplate"><a href="#">Shen Shengqi</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/4/44/Shenshengqi2.JPG" alt="Shen Shengqi" align="left"><p>Everyone considers that it is honored to be a friend of "Brother Face" as he is a totally "local tyrant". Actually," Brother Shen" is warmth, nice, really expert in digging shortcuts in the experiments. He is a sharp soldier of our team as he adheres to the "more with less" principle.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/6/60/Shaoxueying1.JPG" alt="Shao Xueying"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/e/e9/Shaoxueying2.JPG" alt="Shao Xueying"><br />
<div class ="nameplate"><a href="#">Shao Xueying</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/e/e9/Shaoxueying2.JPG" alt="Shao Xueying" align="left"><p>Competent and independent, Shao Xueying has unique ideas about colors and graphics. At the same time, she is a good lecturer. She edits our wiki and does presentation for us. </p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/f/f9/Yuanye1.JPG" alt="Yvette Yuan"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/8/81/Yuanye2.JPG" alt="Yvette Yuan" ><br />
<div class ="nameplate"><a href="#">Yvette Yuan</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/8/81/Yuanye2.JPG" alt="Yvette Yuan" align="left"><p>As a s pronoun for efficient, Yuan Ye is studious and decisive. And what makes her best is that she always brings us delicious oranges.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/f/f3/Yanggege1.JPG" alt="Gege Yang"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/2/25/Yanggege2.JPG" alt="Gege Yang"><br />
<div class ="nameplate"><a href="#">Gege Yang</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/2/25/Yanggege2.JPG" alt="Gege Yang" align="left"><p>She is a lovely girl that everyone likes her, she is also a good experimenter that every step of our project has her contribution.</p><br />
</div><br />
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<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/2/23/Limingyue1.JPG" alt="Mingyue Li"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/0/05/Limingyue2.JPG" alt="Mingyue Li"><br />
<div class ="nameplate"><a href="#">Mingyue Li</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/0/05/Limingyue2.JPG" alt="Mingyue Li" align="left"><p>"Mingyue Bacteria" is the spokesperson of our bacterium, handling the destiny of those little lives. We all agreed that, Mingyue with rubber gloves is GORGEOUSNESS!</p></div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/c/c1/Madanyi1.JPG" alt="Danyi Ma"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/1/19/Madanyi2.JPG" alt="Danyi Ma" ><br />
<div class ="nameplate"><a href="#">Danyi Ma</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/1/19/Madanyi2.JPG" alt="Danyi Ma" align="left"><p>"Aunty Ma" takes charge of our finance and safety, which calls for much patience and responsibility. In the experiment, she also plays an absolutely necessary role.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/7/76/Panminghao1.JPG" alt="Minghao Pan"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/e/ed/Panminghao2.JPG" alt="Minghao Pan" ><br />
<div class ="nameplate"><a href="#">Minghao Pan</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/e/ed/Panminghao2.JPG" alt="Minghao Pan" align="left"><p>Modeling, modeling, and modeling. PS: Taking photos is really not my cup of tea.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/8/87/Zhangheng1.JPG" alt="Zhang Heng"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/6/63/Zhangheng2.JPG" alt="Zhang Heng" ><br />
<div class ="nameplate"><a href="#">Zhang Heng</a> </div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/6/63/Zhangheng2.JPG" alt="Zhang Heng" align="left"><p>He is a man full of responsibility, we could 100% trust him! Bro, it's you that bring us positive energy!</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/1/18/Chenzhuo1.JPG" alt="Chen Zhuo"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/a/af/Chenzhuo2.JPG" alt="Chen Zhuo"><br />
<div class ="nameplate"><a href="#">Chen Zhuo</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/a/af/Chenzhuo2.JPG" alt="Chen Zhuo" align="left"><p>Always, he is still of tongue, but he is not only a genius of experiment, but also a brilliant living library. I cannot say more but admire! </p><br />
</div><br />
</div><br />
</div><br />
<div class ="row"><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/1/15/Wuming1.JPG" alt="Min Wu"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/b/be/Wuming2.JPG" alt="Min Wu"><br />
<div class ="nameplate"><a href="#">Min Wu</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/b/be/Wuming2.JPG" alt="Min Wu" align="left"><p>I love experiments. I love games. I love Weibo.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/d/df/Pengyali1.JPG" alt="Yali Peng"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/0/0c/Pengyali2.JPG" alt="Yali Peng"><br />
<div class ="nameplate"><a href="#">Yali Peng</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/0/0c/Pengyali2.JPG" alt="Yali Peng" align="left"><p>She is a quiet girl, she likes smile, she loves doing experiments peacefully and slowly. As the best partner of Mingyue Li, every trouble become easy! Hey, little pretty we all love you! </p><br />
</div><br />
</div> <br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/3/3f/Chenzhaoxiong1.JPG" alt="Zhaoxiong Chen"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/d/dc/Chenzhaoxiong2.JPG" alt="Zhaoxiong Chen"><br />
<div class ="nameplate"><a href="#">Zhaoxiong Chen</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/d/dc/Chenzhaoxiong2.JPG" alt="Zhaoxiong Chen" align="left"><p>This smart boy is good at playing all kinds of computer systems. We believe that he will refresh the history of wikis!</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/4/4a/Xuehao1.JPG" alt="Hao Xue"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/0/08/Xuehao2.JPG" alt="Hao Xue"><br />
<div class ="nameplate"><a href="#">Hao Xue</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/0/08/Xuehao2.JPG" alt="Hao Xue" align="left"><p>Laughing, he is still laughing! What on hill? Oh god, negative results, but how… But bro, thank you for giving us positive energy, you really raise us up! My bro! </p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/4/49/Qiuyanning1.JPG" alt="Yanning Qiu"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/d/df/Qiuyanning2.JPG" alt="Yanning Qiu"><br />
<div class ="nameplate"><a href="#">Yanning Qiu</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/d/df/Qiuyanning2.JPG" alt="Yanning Qiu" align="left"><p>Our little sister holds the trump cards. She always knows what do with all the words and pictures. Brave and creative, Yanning enjoys the days with new skills and knowledge. Our team was painted colorfully with the lively girl.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/5/52/Longjie1.JPG" alt="Long jie"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/9/96/Longjie2.JPG" alt="Long jie"><br />
<div class ="nameplate"><a href="#">Long jie</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/9/96/Longjie2.JPG" alt="Long jie" align="left"><p>Clever and hard-working, I cannot agree more to do experiments with him. You never let us down!</p><br />
</div><br />
</div><br />
</div><br />
<div class ="row"><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/c/c8/Dongbo1.JPG" alt="Bo Dong"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/0/08/Dongbo2.JPG" alt="Bo Dong" ><br />
<div class ="nameplate"><a href="#">Bo Dong</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/0/08/Dongbo2.JPG" alt="Bo Dong" align="left"><p>He is an earnest boy, he always work hard that every bros and sis like him, he is our team's MVP! <br />
Hey, bro! It our pleasure to be with you!</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/1/14/Wangshiwei1.JPG" alt="Shiwei Wang"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/e/e2/Wangshiwei2.JPG" alt="Shiwei Wang" ><br />
<div class ="nameplate"><a href="#">Shiwei Wang</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/e/e2/Wangshiwei2.JPG" alt="Shiwei Wang" align="left"><p>Again, a quiet boy is coming! He love experiment, he is Bo Dong's loyal friend. We all believe in him, without his help we cannot achieve our goal! Thanks a lot ,my bro! </p><br />
</div><br />
</div> <br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/a/a3/Xinghuayue1.JPG" alt="Huayue Xing"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/a/ae/Xinghuayue2.JPG" alt="Huayue Xing" ><br />
<div class ="nameplate"><a href="#">Huayue Xing</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/a/ae/Xinghuayue2.JPG" alt="Huayue Xing" align="left"><p>With my little eyes, I see bacterium; with my little eyes, I see TD-1; with my little eyes, I see vaccine secreted out; with my little eyes, I see the future without needles. Carefulness, earnest, and a little bit of acting cute, I am XHY.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/a/ad/Xionglei1.JPG" alt="Lei Xiong"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/b/b2/Xionglei2.JPG" alt="Lei Xiong" ><br />
<div class ="nameplate"><a href="#">Lei Xiong</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/b/b2/Xionglei2.JPG" alt="Lei Xiong" align="left"><p>Despite the fact that it is me who always breaks test tubes, loses beakers, and pours reagent onto the skin of my hands, I have the enthusiasm for science. I love to explore and pursue knowledge. As long as there is a chance to see the tip of the iceberg, it doesn't matter how many test tubes I am going to break.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/b/bd/Caoqinjingwen1.JPG" alt="Cao Qinjingwen"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/f/f9/Caoqinjingwen2.JPG" alt="Cao Qinjingwen"><br />
<div class ="nameplate"><a href="#">Cao Qinjingwen</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/f/f9/Caoqinjingwen2.JPG" alt="Cao Qinjingwen" align="left"><p>Excellent! Without these kinds of words, how can I say anything to describe her? As our elder sister, she always gives us self-confident, hey soul sister!</p><br />
</div><br />
</div> <br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/4/4a/Xiaozhuyun1.JPG" alt="Xiao Zhuyun"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/1/1d/Xiaozhuyun2.JPG" alt="Xiao Zhuyun"><br />
<div class ="nameplate"><a href="#">Xiao Zhuyun</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/1/1d/Xiaozhuyun2.JPG" alt="Xiao Zhuyun" align="left"><p>Sincere and straightforward, "piggy", the curve wrecker in our eyes, puts all her efforts into research and study. Only when you get close to her, will you find that she also loves to play, that she also loves life.</p><br />
</div><br />
</div> <br />
</div><br />
<div class ="row"><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/9/9a/Wangzeyu1.JPG" alt="Zeyu Wang"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/5/53/Wangzeyu2.JPG" alt="Zeyu Wang"><br />
<div class ="nameplate"><a href="#">Zeyu Wang</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/5/53/Wangzeyu2.JPG" alt="Zeyu Wang" align="left"><p>I love modeling, too. I hate posing for photographs, neither.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/6/62/Hanyingying1.JPG" alt="Han Yingying"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/c/ca/Hanyingying2.JPG" alt="Han Yingying"><br />
<div class ="nameplate"><a href="#">Han Yingying</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/c/ca/Hanyingying2.JPG" alt="Han Yingying" align="left"><p>Tender as a new-born kitty, Yingying doesn't like to stand in the spotlight. She's a diligent brain instead of a silken tongue. We believe that gold will shine no matter where it is. </p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/e/e4/Fansijia1.JPG" alt="Sijia Fan"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/5/57/Fansijia2" alt="Sijia Fan"><br />
<div class ="nameplate"><a href="#">Sijia Fan</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/5/57/Fansijia2" alt="Sijia Fan" align="left"><p>Black humorist, and sadly, the leader is always shouting at me:" Hurry! Hurry!".</p><br />
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<div class ="row"><br />
<h2>Advisers</h2> <br />
<div class ="profilewrap" style="width:160px;margin:60px 80px 5px;"><br />
<img style="width:160px" src="https://static.igem.org/mediawiki/2013/f/f0/Haiyan_Liu.jpg" alt="Haiyan Liu"><br />
<div class ="nameplate"><a href="#">Haiyan Liu</a></div><br />
<div class = "details"><img style="width:160px" src="https://static.igem.org/mediawiki/2013/f/f0/Haiyan_Liu.jpg" alt="Haiyan Liu" align="left"><p>Haiyan Liu was born in Sichuan Province, China. He received his BS degree in Biology in 1990 and PhD degree in Biochemistry and Molecular Biology in 1996, both from USTC. Between 1993 and 1995 he was a visiting graduate student in Laboratory of Physical Chemistry of ETH, Zurich (Switzerland). Since 2001, he has been a professor of computational biology at School of Life Sciences, USTC. </p><br />
</div><br />
</div><br />
<div class ="profilewrap" style="width:160px;margin:60px 80px 5px;"><br />
<img style="width:160px" src="https://static.igem.org/mediawiki/2013/0/09/Hongjiong.PNG" alt="Jiong Hong"><br />
<div class ="nameplate"><a href="#">Jiong Hong</a></div><br />
<div class = "details"><img style="width:160px" src="https://static.igem.org/mediawiki/2013/0/09/Hongjiong.PNG" alt="Jiong Hong" align="left"><p> I am applying this strategy on the mechanism of the complex diseases such as cancer and diabetes. My ongoing project is to identify biomarkers in order to detect the progress stages of the diabetes. In addition, I have planed to analyze the genetic and environmental factors and their interactions during the progressing of the type 2 diabetes with systems-biology approaches.</p><br />
</div><br />
</div><br />
<div class ="profilewrap" style="width:160px;margin:60px 80px 5px;"><br />
<img style="width:160px" src="https://static.igem.org/mediawiki/2013/4/46/Wujiarui.PNG" alt="Jiarui Wu"><br />
<div class ="nameplate"><a href="#">Jiarui Wu</a></div><br />
<div class = "details"><img style="width:160px" src="https://static.igem.org/mediawiki/2013/4/46/Wujiarui.PNG" alt="Jiarui Wu" align="left"><p>Since the research strategy of systems biology is well fit to analyze the biological complex systems. I am applying this strategy on the mechanism of the complex diseases such as cancer and diabetes. We have developed systematic approaches based on proteomics and bioinformatics to analyze human normal and diabetic serum.</p><br />
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</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/TeamTeam:USTC CHINA/Team2013-09-27T23:29:31Z<p>Surine: </p>
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li><br />
</ul><br />
<div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" /><br />
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<h2>Students</h2><br />
<div class ="row"><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/2/21/Zhangsitao1.JPG" alt="Zhang Sitao"> <br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/0/0d/Zhangsitao2.JPG" alt="Zhang Sitao"><br />
<div class ="nameplate"><a href="#">Zhang Sitao</a> </div><br />
<div class = "details"><br />
<img src="https://static.igem.org/mediawiki/2013/0/0d/Zhangsitao2.JPG" alt="Zhang Sitao" align="left"><p>Our labor leader weighs various matters, leads the overall trend and plays our cards right. He leaves a strong impression in others' mind. However, His friends found that the leader is very cute.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/0/02/Zhaochanglong1.JPG" alt="Changlong Zhao"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/7/7e/Zhaochanglong2.JPG" alt="Changlong Zhao" ><br />
<div class ="nameplate"><a href="#">Changlong Zhao</a></div><br />
<div class = "details"><br />
<img src="https://static.igem.org/mediawiki/2013/7/7e/Zhaochanglong2.JPG" alt="Changlong Zhao" align="left"><p>There's no doubt that we can give full stars for Changlong's fighting capacity. The roads leading to success will never be smooth and Changlong is a perfect companion to travel with.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/9/91/Xionghanjin1.JPG" alt="Hanjin Xiong"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/d/d9/Xionghanjin2.JPG" alt="Hanjin Xiong" ><br />
<div class ="nameplate"><a href="#">Hanjin Xiong</a></div><br />
<div class = "details"><br />
<img src="https://static.igem.org/mediawiki/2013/d/d9/Xionghanjin2.JPG" alt="Hanjin Xiong" align="left"><p>As the keynote speaker of our team, he always keeps a clear head with extraordinary creativity and expressiveness. He said, "It is shameful if you haven't burnt the midnight oil for iGEM." Moving forward bravely, he shows us overwhelming power which nobody can stop it.</p><br />
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</div><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/8/85/Shenshengqi1.JPG" alt="Shen Shengqi"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/4/44/Shenshengqi2.JPG" alt="Shen Shengqi" ><br />
<div class ="nameplate"><a href="#">Shen Shengqi</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/4/44/Shenshengqi2.JPG" alt="Shen Shengqi" align="left"><p>Everyone considers that it is honored to be a friend of "Brother Face" as he is a totally "local tyrant". Actually," Brother Shen" is warmth, nice, really expert in digging shortcuts in the experiments. He is a sharp soldier of our team as he adheres to the "more with less" principle.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/6/60/Shaoxueying1.JPG" alt="Shao Xueying"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/e/e9/Shaoxueying2.JPG" alt="Shao Xueying"><br />
<div class ="nameplate"><a href="#">Shao Xueying</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/e/e9/Shaoxueying2.JPG" alt="Shao Xueying" align="left"><p>Competent and independent, Shao Xueying has unique ideas about colors and graphics. At the same time, she is a good lecturer. She edits our wiki and does presentation for us. </p><br />
</div><br />
</div><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/f/f9/Yuanye1.JPG" alt="Yvette Yuan"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/8/81/Yuanye2.JPG" alt="Yvette Yuan" ><br />
<div class ="nameplate"><a href="#">Yvette Yuan</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/8/81/Yuanye2.JPG" alt="Yvette Yuan" align="left"><p>As a s pronoun for efficient, Yuan Ye is studious and decisive. And what makes her best is that she always brings us delicious oranges.</p><br />
</div><br />
</div><br />
</div><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/f/f3/Yanggege1.JPG" alt="Gege Yang"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/2/25/Yanggege2.JPG" alt="Gege Yang"><br />
<div class ="nameplate"><a href="#">Gege Yang</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/2/25/Yanggege2.JPG" alt="Gege Yang" align="left"><p>She is a lovely girl that everyone likes her, she is also a good experimenter that every step of our project has her contribution.</p><br />
</div><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/2/23/Limingyue1.JPG" alt="Mingyue Li"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/0/05/Limingyue2.JPG" alt="Mingyue Li"><br />
<div class ="nameplate"><a href="#">Mingyue Li</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/0/05/Limingyue2.JPG" alt="Mingyue Li" align="left"><p>"Mingyue Bacteria" is the spokesperson of our bacterium, handling the destiny of those little lives. We all agreed that, Mingyue with rubber gloves is GORGEOUSNESS!</p></div><br />
</div><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/c/c1/Madanyi1.JPG" alt="Danyi Ma"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/1/19/Madanyi2.JPG" alt="Danyi Ma" ><br />
<div class ="nameplate"><a href="#">Danyi Ma</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/1/19/Madanyi2.JPG" alt="Danyi Ma" align="left"><p>"Aunty Ma" takes charge of our finance and safety, which calls for much patience and responsibility. In the experiment, she also plays an absolutely necessary role.</p><br />
</div><br />
</div><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/7/76/Panminghao1.JPG" alt="Minghao Pan"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/e/ed/Panminghao2.JPG" alt="Minghao Pan" ><br />
<div class ="nameplate"><a href="#">Minghao Pan</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/e/ed/Panminghao2.JPG" alt="Minghao Pan" align="left"><p>Modeling, modeling, and modeling. PS: Taking photos is really not my cup of tea.</p><br />
</div><br />
</div><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/8/87/Zhangheng1.JPG" alt="Zhang Heng"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/6/63/Zhangheng2.JPG" alt="Zhang Heng" ><br />
<div class ="nameplate"><a href="#">Zhang Heng</a> </div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/6/63/Zhangheng2.JPG" alt="Zhang Heng" align="left"><p>He is a man full of responsibility, we could 100% trust him! Bro, it's you that bring us positive energy!</p><br />
</div><br />
</div><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/1/18/Chenzhuo1.JPG" alt="Chen Zhuo"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/a/af/Chenzhuo2.JPG" alt="Chen Zhuo"><br />
<div class ="nameplate"><a href="#">Chen Zhuo</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/a/af/Chenzhuo2.JPG" alt="Chen Zhuo" align="left"><p>Always, he is still of tongue, but he is not only a genius of experiment, but also a brilliant living library. I cannot say more but admire! </p><br />
</div><br />
</div><br />
</div><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/2013/1/15/Wuming1.JPG" alt="Min Wu"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/b/be/Wuming2.JPG" alt="Min Wu"><br />
<div class ="nameplate"><a href="#">Min Wu</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/b/be/Wuming2.JPG" alt="Min Wu" align="left"><p>I love experiments. I love games. I love Weibo.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/d/df/Pengyali1.JPG" alt="Yali Peng"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/0/0c/Pengyali2.JPG" alt="Yali Peng"><br />
<div class ="nameplate"><a href="#">Yali Peng</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/0/0c/Pengyali2.JPG" alt="Yali Peng" align="left"><p>She is a quiet girl, she likes smile, she loves doing experiments peacefully and slowly. As the best partner of Mingyue Li, every trouble become easy! Hey, little pretty we all love you! </p><br />
</div><br />
</div> <br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/3/3f/Chenzhaoxiong1.JPG" alt="Zhaoxiong Chen"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/d/dc/Chenzhaoxiong2.JPG" alt="Zhaoxiong Chen"><br />
<div class ="nameplate"><a href="#">Zhaoxiong Chen</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/d/dc/Chenzhaoxiong2.JPG" alt="Zhaoxiong Chen" align="left"><p>This smart boy is good at playing all kinds of computer systems. We believe that he will refresh the history of wikis!</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/4/4a/Xuehao1.JPG" alt="Hao Xue"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/0/08/Xuehao2.JPG" alt="Hao Xue"><br />
<div class ="nameplate"><a href="#">Hao Xue</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/0/08/Xuehao2.JPG" alt="Hao Xue" align="left"><p>Laughing, he is still laughing! What on hill? Oh god, negative results, but how… But bro, thank you for giving us positive energy, you really raise us up! My bro! </p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/4/49/Qiuyanning1.JPG" alt="Yanning Qiu"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/d/df/Qiuyanning2.JPG" alt="Yanning Qiu"><br />
<div class ="nameplate"><a href="#">Yanning Qiu</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/d/df/Qiuyanning2.JPG" alt="Yanning Qiu" align="left"><p>Our little sister holds the trump cards. She always knows what do with all the words and pictures. Brave and creative, Yanning enjoys the days with new skills and knowledge. Our team was painted colorfully with the lively girl.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/5/52/Longjie1.JPG" alt="Long jie"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/9/96/Longjie2.JPG" alt="Long jie"><br />
<div class ="nameplate"><a href="#">Long jie</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/9/96/Longjie2.JPG" alt="Long jie" align="left"><p>Clever and hard-working, I cannot agree more to do experiments with him. You never let us down!</p><br />
</div><br />
</div><br />
</div><br />
<div class ="row"><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/c/c8/Dongbo1.JPG" alt="Bo Dong"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/0/08/Dongbo2.JPG" alt="Bo Dong" ><br />
<div class ="nameplate"><a href="#">Bo Dong</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/0/08/Dongbo2.JPG" alt="Bo Dong" align="left"><p>He is an earnest boy, he always work hard that every bros and sis like him, he is our team's MVP! <br />
Hey, bro! It our pleasure to be with you!</p><br />
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<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/1/14/Wangshiwei1.JPG" alt="Shiwei Wang"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/e/e2/Wangshiwei2.JPG" alt="Shiwei Wang" ><br />
<div class ="nameplate"><a href="#">Shiwei Wang</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/e/e2/Wangshiwei2.JPG" alt="Shiwei Wang" align="left"><p>Again, a quiet boy is coming! He love experiment, he is Bo Dong's loyal friend. We all believe in him, without his help we cannot achieve our goal! Thanks a lot ,my bro! </p><br />
</div><br />
</div> <br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/a/a3/Xinghuayue1.JPG" alt="Huayue Xing"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/a/ae/Xinghuayue2.JPG" alt="Huayue Xing" ><br />
<div class ="nameplate"><a href="#">Huayue Xing</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/a/ae/Xinghuayue2.JPG" alt="Huayue Xing" align="left"><p>With my little eyes, I see bacterium; with my little eyes, I see TD-1; with my little eyes, I see vaccine secreted out; with my little eyes, I see the future without needles. Carefulness, earnest, and a little bit of acting cute, I am XHY.</p><br />
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<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/a/ad/Xionglei1.JPG" alt="Lei Xiong"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/b/b2/Xionglei2.JPG" alt="Lei Xiong" ><br />
<div class ="nameplate"><a href="#">Lei Xiong</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/b/b2/Xionglei2.JPG" alt="Lei Xiong" align="left"><p>Despite the fact that it is me who always breaks test tubes, loses beakers, and pours reagent onto the skin of my hands, I have the enthusiasm for science. I love to explore and pursue knowledge. As long as there is a chance to see the tip of the iceberg, it doesn't matter how many test tubes I am going to break.</p><br />
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<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/b/bd/Caoqinjingwen1.JPG" alt="Cao Qinjingwen"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/f/f9/Caoqinjingwen2.JPG" alt="Cao Qinjingwen"><br />
<div class ="nameplate"><a href="#">Cao Qinjingwen</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/f/f9/Caoqinjingwen2.JPG" alt="Cao Qinjingwen" align="left"><p>Excellent! Without these kinds of words, how can I say anything to describe her? As our elder sister, she always gives us self-confident, hey soul sister!</p><br />
</div><br />
</div> <br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/4/4a/Xiaozhuyun1.JPG" alt="Xiao Zhuyun"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/1/1d/Xiaozhuyun2.JPG" alt="Xiao Zhuyun"><br />
<div class ="nameplate"><a href="#">Xiao Zhuyun</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/1/1d/Xiaozhuyun2.JPG" alt="Xiao Zhuyun" align="left"><p>Sincere and straightforward, "piggy", the curve wrecker in our eyes, puts all her efforts into research and study. Only when you get close to her, will you find that she also loves to play, that she also loves life.</p><br />
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</div> <br />
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<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/9/9a/Wangzeyu1.JPG" alt="Zeyu Wang"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/5/53/Wangzeyu2.JPG" alt="Zeyu Wang"><br />
<div class ="nameplate"><a href="#">Zeyu Wang</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/5/53/Wangzeyu2.JPG" alt="Zeyu Wang" align="left"><p>I love modeling, too. I hate posing for photographs, neither.</p><br />
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<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/6/62/Hanyingying1.JPG" alt="Han Yingying"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/c/ca/Hanyingying2.JPG" alt="Han Yingying"><br />
<div class ="nameplate"><a href="#">Han Yingying</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/c/ca/Hanyingying2.JPG" alt="Han Yingying" align="left"><p>Tender as a new-born kitty, Yingying doesn't like to stand in the spotlight. She's a diligent brain instead of a silken tongue. We believe that gold will shine no matter where it is. </p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/e/e4/Fansijia1.JPG" alt="Sijia Fan"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/5/57/Fansijia2" alt="Sijia Fan"><br />
<div class ="nameplate"><a href="#">Sijia Fan</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/5/57/Fansijia2" alt="Sijia Fan" align="left"><p>Black humorist, and sadly, the leader is always shouting at me:" Hurry! Hurry!".</p><br />
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<h2>Advisers</h2> <br />
<div class ="profilewrap" style="width:160px;margin:60px 80px 5px;"><br />
<img style="width:160px" src="https://static.igem.org/mediawiki/2013/f/f0/Haiyan_Liu.jpg" alt="Haiyan Liu"><br />
<div class ="nameplate"><a href="#">Haiyan Liu</a></div><br />
<div class = "details"><img style="width:160px" src="https://static.igem.org/mediawiki/2013/f/f0/Haiyan_Liu.jpg" alt="Haiyan Liu" align="left"><p>Haiyan Liu was born in Sichuan Province, China. He received his BS degree in Biology in 1990 and PhD degree in Biochemistry and Molecular Biology in 1996, both from USTC. Between 1993 and 1995 he was a visiting graduate student in Laboratory of Physical Chemistry of ETH, Zurich (Switzerland). Since 2001, he has been a professor of computational biology at School of Life Sciences, USTC. </p><br />
</div><br />
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<div class ="profilewrap" style="width:160px;margin:60px 80px 5px;"><br />
<img style="width:160px" src="https://static.igem.org/mediawiki/2013/0/09/Hongjiong.PNG" alt="Jiong Hong"><br />
<div class ="nameplate"><a href="#">Jiong Hong</a></div><br />
<div class = "details"><img style="width:160px" src="https://static.igem.org/mediawiki/2013/0/09/Hongjiong.PNG" alt="Jiong Hong" align="left"><p> I am applying this strategy on the mechanism of the complex diseases such as cancer and diabetes. My ongoing project is to identify biomarkers in order to detect the progress stages of the diabetes. In addition, I have planed to analyze the genetic and environmental factors and their interactions during the progressing of the type 2 diabetes with systems-biology approaches.</p><br />
</div><br />
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<div class ="profilewrap" style="width:160px;margin:60px 80px 5px;"><br />
<img style="width:160px" src="https://static.igem.org/mediawiki/2013/4/46/Wujiarui.PNG" alt="Jiarui Wu"><br />
<div class ="nameplate"><a href="#">Jiarui Wu</a></div><br />
<div class = "details"><img style="width:160px" src="https://static.igem.org/mediawiki/2013/4/46/Wujiarui.PNG" alt="Jiarui Wu" align="left"><p>Since the research strategy of systems biology is well fit to analyze the biological complex systems. I am applying this strategy on the mechanism of the complex diseases such as cancer and diabetes. We have developed systematic approaches based on proteomics and bioinformatics to analyze human normal and diabetic serum.</p><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Background">Background</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Design">Design</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></li><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">Notebook</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">Timeline</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li><br />
</ul><br />
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<li class="active"><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Members</a></li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=USTC_CHINA">Profile</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Attributions">Attributions</a></li><br />
<li><a href="https://igem.org/2013_Judging_Form?id=1074#iGEM_Medals">Achievements</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li><br />
</ul><br />
<div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" /><br />
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<div class="content" align="center" style="margin-bottom: 50px"><br />
<h2>Students</h2><br />
<div class ="row"><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/2/21/Zhangsitao1.JPG" alt="Zhang Sitao"> <br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/0/0d/Zhangsitao2.JPG" alt="Zhang Sitao"><br />
<div class ="nameplate"><a href="#">Zhang Sitao</a> </div><br />
<div class = "details"><br />
<img src="https://static.igem.org/mediawiki/2013/0/0d/Zhangsitao2.JPG" alt="Zhang Sitao" align="left"><p>Our labor leader weighs various matters, leads the overall trend and plays our cards right. He leaves a strong impression in others' mind. However, His friends found that the leader is very cute.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/0/02/Zhaochanglong1.JPG" alt="Changlong Zhao"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/7/7e/Zhaochanglong2.JPG" alt="Changlong Zhao" ><br />
<div class ="nameplate"><a href="#">Changlong Zhao</a></div><br />
<div class = "details"><br />
<img src="https://static.igem.org/mediawiki/2013/7/7e/Zhaochanglong2.JPG" alt="Changlong Zhao" align="left"><p>There's no doubt that we can give full stars for Changlong's fighting capacity. The roads leading to success will never be smooth and Changlong is a perfect companion to travel with.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/9/91/Xionghanjin1.JPG" alt="Hanjin Xiong"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/d/d9/Xionghanjin2.JPG" alt="Hanjin Xiong" ><br />
<div class ="nameplate"><a href="#">Hanjin Xiong</a></div><br />
<div class = "details"><br />
<img src="https://static.igem.org/mediawiki/2013/d/d9/Xionghanjin2.JPG" alt="Hanjin Xiong" align="left"><p>As the keynote speaker of our team, he always keeps a clear head with extraordinary creativity and expressiveness. He said, "It is shameful if you haven't burnt the midnight oil for iGEM." Moving forward bravely, he shows us overwhelming power which nobody can stop it.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/8/85/Shenshengqi1.JPG" alt="Shen Shengqi"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/4/44/Shenshengqi2.JPG" alt="Shen Shengqi" ><br />
<div class ="nameplate"><a href="#">Shen Shengqi</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/4/44/Shenshengqi2.JPG" alt="Shen Shengqi" align="left"><p>Everyone considers that it is honored to be a friend of "Brother Face" as he is a totally "local tyrant". Actually," Brother Shen" is warmth, nice, really expert in digging shortcuts in the experiments. He is a sharp soldier of our team as he adheres to the "more with less" principle.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/6/60/Shaoxueying1.JPG" alt="Shao Xueying"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/e/e9/Shaoxueying2.JPG" alt="Shao Xueying"><br />
<div class ="nameplate"><a href="#">Shao Xueying</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/e/e9/Shaoxueying2.JPG" alt="Shao Xueying" align="left"><p>Competent and independent, Shao Xueying has unique ideas about colors and graphics. At the same time, she is a good lecturer. She edits our wiki and does presentation for us. </p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/f/f9/Yuanye1.JPG" alt="Yvette Yuan"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/8/81/Yuanye2.JPG" alt="Yvette Yuan" ><br />
<div class ="nameplate"><a href="#">Yvette Yuan</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/8/81/Yuanye2.JPG" alt="Yvette Yuan" align="left"><p>As a s pronoun for efficient, Yuan Ye is studious and decisive. And what makes her best is that she always brings us delicious oranges.</p><br />
</div><br />
</div><br />
</div><br />
<div class ="row"><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/f/f3/Yanggege1.JPG" alt="Gege Yang"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/2/25/Yanggege2.JPG" alt="Gege Yang"><br />
<div class ="nameplate"><a href="#">Gege Yang</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/2/25/Yanggege2.JPG" alt="Gege Yang" align="left"><p>She is a lovely girl that everyone likes her, she is also a good experimenter that every step of our project has her contribution.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/2/23/Limingyue1.JPG" alt="Mingyue Li"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/0/05/Limingyue2.JPG" alt="Mingyue Li"><br />
<div class ="nameplate"><a href="#">Mingyue Li</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/0/05/Limingyue2.JPG" alt="Mingyue Li" align="left"><p>"Mingyue Bacteria" is the spokesperson of our bacterium, handling the destiny of those little lives. We all agreed that, Mingyue with rubber gloves is GORGEOUSNESS!</p></div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/c/c1/Madanyi1.JPG" alt="Danyi Ma"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/1/19/Madanyi2.JPG" alt="Danyi Ma" ><br />
<div class ="nameplate"><a href="#">Danyi Ma</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/1/19/Madanyi2.JPG" alt="Danyi Ma" align="left"><p>"Aunty Ma" takes charge of our finance and safety, which calls for much patience and responsibility. In the experiment, she also plays an absolutely necessary role.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/7/76/Panminghao1.JPG" alt="Minghao Pan"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/e/ed/Panminghao2.JPG" alt="Minghao Pan" ><br />
<div class ="nameplate"><a href="#">Minghao Pan</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/e/ed/Panminghao2.JPG" alt="Minghao Pan" align="left"><p>Modeling, modeling, and modeling. PS: Taking photos is really not my cup of tea.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/8/87/Zhangheng1.JPG" alt="Zhang Heng"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/6/63/Zhangheng2.JPG" alt="Zhang Heng" ><br />
<div class ="nameplate"><a href="#">Zhang Heng</a> </div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/6/63/Zhangheng2.JPG" alt="Zhang Heng" align="left"><p>He is a man full of responsibility, we could 100% trust him! Bro, it's you that bring us positive energy!</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/1/18/Chenzhuo1.JPG" alt="Chen Zhuo"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/a/af/Chenzhuo2.JPG" alt="Chen Zhuo"><br />
<div class ="nameplate"><a href="#">Chen Zhuo</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/a/af/Chenzhuo2.JPG" alt="Chen Zhuo" align="left"><p>Always, he is still of tongue, but he is not only a genius of experiment, but also a brilliant living library. I cannot say more but admire! </p><br />
</div><br />
</div><br />
</div><br />
<div class ="row"><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/1/15/Wuming1.JPG" alt="Min Wu"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/b/be/Wuming2.JPG" alt="Min Wu"><br />
<div class ="nameplate"><a href="#">Min Wu</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/b/be/Wuming2.JPG" alt="Min Wu" align="left"><p>I love experiments. I love games. I love Weibo.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/d/df/Pengyali1.JPG" alt="Yali Peng"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/0/0c/Pengyali2.JPG" alt="Yali Peng"><br />
<div class ="nameplate"><a href="#">Yali Peng</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/0/0c/Pengyali2.JPG" alt="Yali Peng" align="left"><p>She is a quiet girl, she likes smile, she loves doing experiments peacefully and slowly. As the best partner of Mingyue Li, every trouble become easy! Hey, little pretty we all love you! </p><br />
</div><br />
</div> <br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/3/3f/Chenzhaoxiong1.JPG" alt="Zhaoxiong Chen"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/d/dc/Chenzhaoxiong2.JPG" alt="Zhaoxiong Chen"><br />
<div class ="nameplate"><a href="#">Zhaoxiong Chen</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/d/dc/Chenzhaoxiong2.JPG" alt="Zhaoxiong Chen" align="left"><p>This smart boy is good at playing all kinds of computer systems. We believe that he will refresh the history of wikis!</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/4/4a/Xuehao1.JPG" alt="Hao Xue"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/0/08/Xuehao2.JPG" alt="Hao Xue"><br />
<div class ="nameplate"><a href="#">Hao Xue</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/0/08/Xuehao2.JPG" alt="Hao Xue" align="left"><p>Laughing, he is still laughing! What on hill? Oh god, negative results, but how… But bro, thank you for giving us positive energy, you really raise us up! My bro! </p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/4/49/Qiuyanning1.JPG" alt="Yanning Qiu"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/d/df/Qiuyanning2.JPG" alt="Yanning Qiu"><br />
<div class ="nameplate"><a href="#">Yanning Qiu</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/d/df/Qiuyanning2.JPG" alt="Yanning Qiu" align="left"><p>Our little sister holds the trump cards. She always knows what do with all the words and pictures. Brave and creative, Yanning enjoys the days with new skills and knowledge. Our team was painted colorfully with the lively girl.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/5/52/Longjie1.JPG" alt="Long jie"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/9/96/Longjie2.JPG" alt="Long jie"><br />
<div class ="nameplate"><a href="#">Long jie</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/9/96/Longjie2.JPG" alt="Long jie" align="left"><p>Clever and hard-working, I cannot agree more to do experiments with him. You never let us down!</p><br />
</div><br />
</div><br />
</div><br />
<div class ="row"><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/c/c8/Dongbo1.JPG" alt="Bo Dong"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/0/08/Dongbo2.JPG" alt="Bo Dong" ><br />
<div class ="nameplate"><a href="#">Bo Dong</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/0/08/Dongbo2.JPG" alt="Bo Dong" align="left"><p>He is an earnest boy, he always work hard that every bros and sis like him, he is our team's MVP! <br />
Hey, bro! It our pleasure to be with you!</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/1/14/Wangshiwei1.JPG" alt="Shiwei Wang"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/e/e2/Wangshiwei2.JPG" alt="Shiwei Wang" ><br />
<div class ="nameplate"><a href="#">Shiwei Wang</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/e/e2/Wangshiwei2.JPG" alt="Shiwei Wang" align="left"><p>Again, a quiet boy is coming! He love experiment, he is Bo Dong's loyal friend. We all believe in him, without his help we cannot achieve our goal! Thanks a lot ,my bro! </p><br />
</div><br />
</div> <br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/a/a3/Xinghuayue1.JPG" alt="Huayue Xing"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/a/ae/Xinghuayue2.JPG" alt="Huayue Xing" ><br />
<div class ="nameplate"><a href="#">Huayue Xing</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/a/ae/Xinghuayue2.JPG" alt="Huayue Xing" align="left"><p>With my little eyes, I see bacterium; with my little eyes, I see TD-1; with my little eyes, I see vaccine secreted out; with my little eyes, I see the future without needles. Carefulness, earnest, and a little bit of acting cute, I am XHY.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/a/ad/Xionglei1.JPG" alt="Lei Xiong"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/b/b2/Xionglei2.JPG" alt="Lei Xiong" ><br />
<div class ="nameplate"><a href="#">Lei Xiong</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/b/b2/Xionglei2.JPG" alt="Lei Xiong" align="left"><p>Despite the fact that it is me who always breaks test tubes, loses beakers, and pours reagent onto the skin of my hands, I have the enthusiasm for science. I love to explore and pursue knowledge. As long as there is a chance to see the tip of the iceberg, it doesn't matter how many test tubes I am going to break.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/b/bd/Caoqinjingwen1.JPG" alt="Cao Qinjingwen"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/f/f9/Caoqinjingwen2.JPG" alt="Cao Qinjingwen"><br />
<div class ="nameplate"><a href="#">Cao Qinjingwen</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/f/f9/Caoqinjingwen2.JPG" alt="Cao Qinjingwen" align="left"><p>Excellent! Without these kinds of words, how can I say anything to describe her? As our elder sister, she always gives us self-confident, hey soul sister!</p><br />
</div><br />
</div> <br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/4/4a/Xiaozhuyun1.JPG" alt="Xiao Zhuyun"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/1/1d/Xiaozhuyun2.JPG" alt="Xiao Zhuyun"><br />
<div class ="nameplate"><a href="#">Xiao Zhuyun</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/1/1d/Xiaozhuyun2.JPG" alt="Xiao Zhuyun" align="left"><p>Sincere and straightforward, "piggy", the curve wrecker in our eyes, puts all her efforts into research and study. Only when you get close to her, will you find that she also loves to play, that she also loves life.</p><br />
</div><br />
</div> <br />
</div><br />
<div class ="row"><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/9/9a/Wangzeyu1.JPG" alt="Zeyu Wang"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/5/53/Wangzeyu2.JPG" alt="Zeyu Wang"><br />
<div class ="nameplate"><a href="#">Zeyu Wang</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/5/53/Wangzeyu2.JPG" alt="Zeyu Wang" align="left"><p>I love modeling, too. I hate posing for photographs, neither.</p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/igem.org/6/62/Hanyingying1.JPG" alt="Han Yingying"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/igem.org/c/ca/Hanyingying2.JPG" alt="Han Yingying"><br />
<div class ="nameplate"><a href="#">Han Yingying</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/igem.org/c/ca/Hanyingying2.JPG" alt="Han Yingying" align="left"><p>Tender as a new-born kitty, Yingying doesn't like to stand in the spotlight. She's a diligent brain instead of a silken tongue. We believe that gold will shine no matter where it is. </p><br />
</div><br />
</div><br />
<div class ="profilewrap"><br />
<img class ="normal" src="https://static.igem.org/mediawiki/2013/e/e4/Fansijia1.JPG" alt="Sijia Fan"><br />
<img class ="crazy" src="https://static.igem.org/mediawiki/2013/5/57/Fansijia2" alt="Sijia Fan"><br />
<div class ="nameplate"><a href="#">Sijia Fan</a></div><br />
<div class = "details"><img src="https://static.igem.org/mediawiki/2013/5/57/Fansijia2" alt="Sijia Fan" align="left"><p>Black humorist, and sadly, the leader is always shouting at me:" Hurry! Hurry!".</p><br />
</div><br />
</div><br />
</div><br />
<br />
<div class ="row"><br />
<h2>Advisers</h2> <br />
<div class ="profilewrap" style="width:160px;margin:60px 80px 5px;"><br />
<img style="width:160px" src="https://static.igem.org/mediawiki/2013/f/f0/Haiyan_Liu.jpg" alt="Haiyan Liu"><br />
<div class ="nameplate"><a href="#">Haiyan Liu</a></div><br />
<div class = "details"><img style="width:160px" src="https://static.igem.org/mediawiki/2013/f/f0/Haiyan_Liu.jpg" alt="Haiyan Liu" align="left"><p>Haiyan Liu was born in Sichuan Province, China. He received his BS degree in Biology in 1990 and PhD degree in Biochemistry and Molecular Biology in 1996, both from USTC. Between 1993 and 1995 he was a visiting graduate student in Laboratory of Physical Chemistry of ETH, Zurich (Switzerland). Since 2001, he has been a professor of computational biology at School of Life Sciences, USTC. </p><br />
</div><br />
</div><br />
<div class ="profilewrap" style="width:160px;margin:60px 80px 5px;"><br />
<img style="width:160px" src="https://static.igem.org/mediawiki/2013/0/09/Hongjiong.PNG" alt="Jiong Hong"><br />
<div class ="nameplate"><a href="#">Jiong Hong</a></div><br />
<div class = "details"><img style="width:160px" src="https://static.igem.org/mediawiki/2013/0/09/Hongjiong.PNG" alt="Jiong Hong" align="left"><p> I am applying this strategy on the mechanism of the complex diseases such as cancer and diabetes. My ongoing project is to identify biomarkers in order to detect the progress stages of the diabetes. In addition, I have planed to analyze the genetic and environmental factors and their interactions during the progressing of the type 2 diabetes with systems-biology approaches.</p><br />
</div><br />
</div><br />
<div class ="profilewrap" style="width:160px;margin:60px 80px 5px;"><br />
<img style="width:160px" src="https://static.igem.org/mediawiki/2013/4/46/Wujiarui.PNG" alt="Jiarui Wu"><br />
<div class ="nameplate"><a href="#">Jiarui Wu</a></div><br />
<div class = "details"><img style="width:160px" src="https://static.igem.org/mediawiki/2013/4/46/Wujiarui.PNG" alt="Jiarui Wu" align="left"><p>Since the research strategy of systems biology is well fit to analyze the biological complex systems. I am applying this strategy on the mechanism of the complex diseases such as cancer and diabetes. We have developed systematic approaches based on proteomics and bioinformatics to analyze human normal and diabetic serum.</p><br />
</div><br />
</div><br />
</div><br />
</div><br />
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</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/home.cssTeam:USTC CHINA/home.css2013-09-27T22:51:17Z<p>Surine: </p>
<hr />
<div>@charset "utf-8";<br />
/* CSS Document */<br />
#header{<br />
width:960px;<br />
margin-top:10px;<br />
position:relative;<br />
z-index:1900;<br />
}<br />
#logo{<br />
width:190px;<br />
height:123px;<br />
background:url(https://static.igem.org/mediawiki/2013/8/8a/2013ustc-china_Logo-bg.png);<br />
display:block;<br />
float:left;<br />
}<br />
#header #nav{<br />
margin-left:189px;<br />
padding-left:34px;<br />
width:740px;<br />
}<br />
.pic-bar{<br />
width:964px;<br />
height:410px;<br />
margin:10px auto 0 auto;<br />
}<br />
.home-pic{<br />
height:100%;<br />
background:url(https://static.igem.org/mediawiki/2013/d/d4/2013ustc-china_Draft.png) no-repeat scroll left top transparent;<br />
}<br />
.content .pic-bar .home-pic .page-tittle{<br />
margin:0 0 30px;<br />
padding:0 0 5px;<br />
}<br />
.content .pic-bar .home-pic .page-tittle h1{<br />
color: #383636;<br />
font-size: 36px;<br />
font-weight: 500;<br />
text-align: right;<br />
}<br />
.content .pic-bar .home-pic .page-para p{<br />
color: #626364;<br />
font-size: 14px;<br />
line-height: 20px;<br />
padding-bottom:15px;<br />
}<br />
.button-home{<br />
background-color: #393939;<br />
border: 0 none;<br />
cursor: pointer;<br />
display: block;<br />
font-weight: lighter;<br />
margin: 0;<br />
overflow: visible;<br />
padding:0 10px 0 10px;<br />
text-decoration: none;<br />
width: 55px;<br />
}<br />
a.button-home:hover span{<br />
color:#7bc5ff;<br />
}<br />
.page-tittle span{color:#ff9933;}<br />
.button-home span{<br />
color: #FFFFFF;<br />
font: lighter 18px/44px Merriweather,Arial;<br />
height: 44px;<br />
padding: 0;<br />
text-align: center;<br />
white-space: nowrap;<br />
width: 100%;<br />
}<br />
#homecol{<br />
height:1150px;<br />
}<br />
#home.leftbar{height:1150px;}<br />
#home.rightbar{height:1150px;}<br />
#abstract-bar{<br />
margin-top:20px;<br />
margin-bottom:20px;<br />
}<br />
#pro-bar{<br />
width:620px;<br />
height:500px;<br />
}<br />
#homecol .conbar2 .leftbar #abstract-bar #abs-1{<br />
border-radius:1em 1em 1em 1em;<br />
background:#ffffff;<br />
border:1px solid rgb(68,68,68);<br />
margin:10px 0 0 0;<br />
padding:5px 5px 15px 5px;<br />
}<br />
#homecol .conbar2 .leftbar #abstract-bar #abs-1 #abs-text{<br />
text-align:center;<br />
font-size:13px;<br />
line-height:20px;} <br />
<br />
#homecol span{<br />
color:#F93;<br />
font-size:16px;}<br />
#homecol .conbar2 #home #pro-bar #pro-text{<br />
margin:0 0 20px 0;<br />
font-size:14px;<br />
line-height:20px;<br />
}<br />
#homecol .conbar2 #home #pro-bar #pro-de-text{<br />
height:120px;}<br />
.pro-col{<br />
float:left;<br />
width:288px;<br />
height:150px;<br />
margin-bottom:20px;<br />
text-align:center;<br />
border-radius:1em 1em 1em 1em;<br />
background:#ffffff;<br />
border:1px solid rgb(68,68,68);<br />
padding:5px;<br />
}<br />
#ls.pro-col{<br />
margin-right:20px}<br />
#homecol .conbar2 #home #exp-bar .exp-col{<br />
border-radius:1em 1em 1em 1em;<br />
background:#ffffff;<br />
border:1px solid rgb(68,68,68);<br />
text-align:center;<br />
font-size:14px;<br />
line-height:20px;<br />
margin-bottom:10px;<br />
padding:5px 5px 15px 5px;<br />
}<br />
#homecol .conbar2 #home #modle-bar{<br />
margin-top:20px;}<br />
#homecol .conbar2 #home #modle-bar #modle-col{<br />
border-radius:1em 1em 1em 1em;<br />
background:#ffffff;<br />
border:1px solid rgb(68,68,68);<br />
padding:5px;<br />
} <br />
#homecol .conbar2 #home #modle-bar h2{<br />
font-size:16px;<br />
font-weight:100;<br />
margin-top:10px;}<br />
#homecol .conbar2 #home #hum-bar{<br />
margin-top:10px;<br />
}<br />
#homecol .conbar2 #home #hum-bar #hum-col{<br />
border-radius:1em 1em 1em 1em;<br />
background:#ffffff;<br />
border:1px solid rgb(68,68,68);<br />
padding:5px;<br />
}<br />
#homecol .conbar2 #home #hum-bar h2{ <br />
font-size:16px;<br />
font-weight:100;<br />
margin-top:10px;<br />
}<br />
#footer{<br />
color:#fff;<br />
padding-top:30px;<br />
}<br />
#thanks{<br />
width:964px;<br />
height:280px;<br />
margin-top:10px;<br />
margin-left:auto; margin-right:auto;<br />
}<br />
#sponsors-title{<br />
color: #303030;<br />
position:relative;<br />
margin-top:20px;<br />
font-size:20px;<br />
line-height:40px;<br />
}<br />
<br />
.sponsor{<br />
display:block;<br />
float:left;<br />
width:280px;<br />
height:68px;<br />
margin: 20px;<br />
overflow:hidden;<br />
color:#303030;<br />
}<br />
.sponsor:hover p{<br />
margin-top:-65px;<br />
}<br />
.sponsor p{<br />
display:block;<br />
width:290px;<br />
height:65px;<br />
line-height:20px;<br />
font-size:16px;<br />
text-align:center;<br />
vertical-align:center;<br />
margin:0;<br />
<br />
-webkit-transition: margin-top 0.2s linear;<br />
-moz-transition: margin-top 0.2s linear;<br />
-o-transition: margin-top 0.2s linear;<br />
-ms-transition: margin-top 0.2s linear;<br />
transition: margin-top 0.2s linear;<br />
}<br />
.sponsor a img{<br />
opacity:0;<br />
-webkit-transition:all 0.3s linear;<br />
-o-transition:all 0.3s linear;<br />
-ms-transition:all 0.3s linear;<br />
-moz-transition:all 0.3s linear;<br />
transition:all 0.3s linear;<br />
-moz-transform: rotate(6deg);<br />
-o-transform: rotate(6deg);<br />
-webkit-transform: rotate(6deg);<br />
}<br />
.sponsor:hover a img{<br />
opacity:1;<br />
-webkit-transform:rotate(0deg);<br />
-moz-transform:rotate(0deg);<br />
-o-transform:rotate(0deg);<br />
}<br />
<br />
.sponsor a p{<br />
height:45px;<br />
padding:10px;<br />
font-family: 'Petit Formal Script', cursive;<br />
font-size:14px;<br />
<br />
opacity:0;<br />
-webkit-transition:all 0.3s linear;<br />
-o-transition:all 0.3s linear;<br />
-ms-transition:all 0.3s linear;<br />
-moz-transition:all 0.3s linear;<br />
transition:all 0.3s linear;<br />
<br />
-webkit-transform: rotate(6deg);<br />
}<br />
.sponsor:hover a p{<br />
opacity:1;<br />
-webkit-transform:rotate(0deg);<br />
}</div>Surinehttp://2013.igem.org/Team:USTC_CHINA/home.cssTeam:USTC CHINA/home.css2013-09-27T22:50:57Z<p>Surine: </p>
<hr />
<div>@charset "utf-8";<br />
/* CSS Document */<br />
#header{<br />
width:960px;<br />
margin-top:10px;<br />
position:relative;<br />
z-index:1900;<br />
}<br />
#logo{<br />
width:190px;<br />
height:123px;<br />
background:url(https://static.igem.org/mediawiki/2013/8/8a/2013ustc-china_Logo-bg.png);<br />
display:block;<br />
float:left;<br />
}<br />
#header #nav{<br />
margin-left:189px;<br />
padding-left:34px;<br />
width:740px;<br />
}<br />
.pic-bar{<br />
width:964px;<br />
height:410px;<br />
margin:10px auto 0 auto;<br />
}<br />
.home-pic{<br />
height:100%;<br />
background:url(https://static.igem.org/mediawiki/2013/d/d4/2013ustc-china_Draft.png) no-repeat scroll left top transparent;<br />
}<br />
.content .pic-bar .home-pic .page-tittle{<br />
margin:0 0 30px;<br />
padding:0 0 5px;<br />
}<br />
.content .pic-bar .home-pic .page-tittle h1{<br />
color: #383636;<br />
font-size: 36px;<br />
font-weight: 500;<br />
text-align: right;<br />
}<br />
.content .pic-bar .home-pic .page-para p{<br />
color: #626364;<br />
font-size: 14px;<br />
line-height: 20px;<br />
padding-bottom:15px;<br />
}<br />
.button-home{<br />
background-color: #393939;<br />
border: 0 none;<br />
cursor: pointer;<br />
display: block;<br />
font-weight: lighter;<br />
margin: 0;<br />
overflow: visible;<br />
padding:0 10px 0 10px;<br />
text-decoration: none;<br />
width: 55px;<br />
}<br />
a.button-home:hover span{<br />
color:#7bc5ff;<br />
}<br />
.page-tittle span{#ff9933;}<br />
.button-home span{<br />
color: #FFFFFF;<br />
font: lighter 18px/44px Merriweather,Arial;<br />
height: 44px;<br />
padding: 0;<br />
text-align: center;<br />
white-space: nowrap;<br />
width: 100%;<br />
}<br />
#homecol{<br />
height:1150px;<br />
}<br />
#home.leftbar{height:1150px;}<br />
#home.rightbar{height:1150px;}<br />
#abstract-bar{<br />
margin-top:20px;<br />
margin-bottom:20px;<br />
}<br />
#pro-bar{<br />
width:620px;<br />
height:500px;<br />
}<br />
#homecol .conbar2 .leftbar #abstract-bar #abs-1{<br />
border-radius:1em 1em 1em 1em;<br />
background:#ffffff;<br />
border:1px solid rgb(68,68,68);<br />
margin:10px 0 0 0;<br />
padding:5px 5px 15px 5px;<br />
}<br />
#homecol .conbar2 .leftbar #abstract-bar #abs-1 #abs-text{<br />
text-align:center;<br />
font-size:13px;<br />
line-height:20px;} <br />
<br />
#homecol span{<br />
color:#F93;<br />
font-size:16px;}<br />
#homecol .conbar2 #home #pro-bar #pro-text{<br />
margin:0 0 20px 0;<br />
font-size:14px;<br />
line-height:20px;<br />
}<br />
#homecol .conbar2 #home #pro-bar #pro-de-text{<br />
height:120px;}<br />
.pro-col{<br />
float:left;<br />
width:288px;<br />
height:150px;<br />
margin-bottom:20px;<br />
text-align:center;<br />
border-radius:1em 1em 1em 1em;<br />
background:#ffffff;<br />
border:1px solid rgb(68,68,68);<br />
padding:5px;<br />
}<br />
#ls.pro-col{<br />
margin-right:20px}<br />
#homecol .conbar2 #home #exp-bar .exp-col{<br />
border-radius:1em 1em 1em 1em;<br />
background:#ffffff;<br />
border:1px solid rgb(68,68,68);<br />
text-align:center;<br />
font-size:14px;<br />
line-height:20px;<br />
margin-bottom:10px;<br />
padding:5px 5px 15px 5px;<br />
}<br />
#homecol .conbar2 #home #modle-bar{<br />
margin-top:20px;}<br />
#homecol .conbar2 #home #modle-bar #modle-col{<br />
border-radius:1em 1em 1em 1em;<br />
background:#ffffff;<br />
border:1px solid rgb(68,68,68);<br />
padding:5px;<br />
} <br />
#homecol .conbar2 #home #modle-bar h2{<br />
font-size:16px;<br />
font-weight:100;<br />
margin-top:10px;}<br />
#homecol .conbar2 #home #hum-bar{<br />
margin-top:10px;<br />
}<br />
#homecol .conbar2 #home #hum-bar #hum-col{<br />
border-radius:1em 1em 1em 1em;<br />
background:#ffffff;<br />
border:1px solid rgb(68,68,68);<br />
padding:5px;<br />
}<br />
#homecol .conbar2 #home #hum-bar h2{ <br />
font-size:16px;<br />
font-weight:100;<br />
margin-top:10px;<br />
}<br />
#footer{<br />
color:#fff;<br />
padding-top:30px;<br />
}<br />
#thanks{<br />
width:964px;<br />
height:280px;<br />
margin-top:10px;<br />
margin-left:auto; margin-right:auto;<br />
}<br />
#sponsors-title{<br />
color: #303030;<br />
position:relative;<br />
margin-top:20px;<br />
font-size:20px;<br />
line-height:40px;<br />
}<br />
<br />
.sponsor{<br />
display:block;<br />
float:left;<br />
width:280px;<br />
height:68px;<br />
margin: 20px;<br />
overflow:hidden;<br />
color:#303030;<br />
}<br />
.sponsor:hover p{<br />
margin-top:-65px;<br />
}<br />
.sponsor p{<br />
display:block;<br />
width:290px;<br />
height:65px;<br />
line-height:20px;<br />
font-size:16px;<br />
text-align:center;<br />
vertical-align:center;<br />
margin:0;<br />
<br />
-webkit-transition: margin-top 0.2s linear;<br />
-moz-transition: margin-top 0.2s linear;<br />
-o-transition: margin-top 0.2s linear;<br />
-ms-transition: margin-top 0.2s linear;<br />
transition: margin-top 0.2s linear;<br />
}<br />
.sponsor a img{<br />
opacity:0;<br />
-webkit-transition:all 0.3s linear;<br />
-o-transition:all 0.3s linear;<br />
-ms-transition:all 0.3s linear;<br />
-moz-transition:all 0.3s linear;<br />
transition:all 0.3s linear;<br />
-moz-transform: rotate(6deg);<br />
-o-transform: rotate(6deg);<br />
-webkit-transform: rotate(6deg);<br />
}<br />
.sponsor:hover a img{<br />
opacity:1;<br />
-webkit-transform:rotate(0deg);<br />
-moz-transform:rotate(0deg);<br />
-o-transform:rotate(0deg);<br />
}<br />
<br />
.sponsor a p{<br />
height:45px;<br />
padding:10px;<br />
font-family: 'Petit Formal Script', cursive;<br />
font-size:14px;<br />
<br />
opacity:0;<br />
-webkit-transition:all 0.3s linear;<br />
-o-transition:all 0.3s linear;<br />
-ms-transition:all 0.3s linear;<br />
-moz-transition:all 0.3s linear;<br />
transition:all 0.3s linear;<br />
<br />
-webkit-transform: rotate(6deg);<br />
}<br />
.sponsor:hover a p{<br />
opacity:1;<br />
-webkit-transform:rotate(0deg);<br />
}</div>Surinehttp://2013.igem.org/Team:USTC_CHINA/home.cssTeam:USTC CHINA/home.css2013-09-27T22:49:03Z<p>Surine: </p>
<hr />
<div>@charset "utf-8";<br />
/* CSS Document */<br />
#header{<br />
width:960px;<br />
margin-top:10px;<br />
position:relative;<br />
z-index:1900;<br />
}<br />
#logo{<br />
width:190px;<br />
height:123px;<br />
background:url(https://static.igem.org/mediawiki/2013/8/8a/2013ustc-china_Logo-bg.png);<br />
display:block;<br />
float:left;<br />
}<br />
#header #nav{<br />
margin-left:189px;<br />
padding-left:34px;<br />
width:740px;<br />
}<br />
.pic-bar{<br />
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}</div>Surinehttp://2013.igem.org/Team:USTC_CHINATeam:USTC CHINA2013-09-27T22:43:53Z<p>Surine: </p>
<hr />
<div>{{USTC-China/hidden}}<br />
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<div id="header"><br />
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<img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="190" height="123" /><br />
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<ul id="nav"><br />
<li class="active"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Background">Background</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Design">Design</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">Notebook</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">Timeline</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Members</a></li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=USTC_CHINA">Profile</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Attributions">Attributions</a></li><br />
<li><a href="https://igem.org/2013_Judging_Form?id=1074#iGEM_Medals">Achievements</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li><br />
</ul><br />
<div id="igemlogo" style="float:right;margin: -48px 20px 0px 0px;"><br />
<a href="https://2013.igem.org/Main_Page" target="_blank"><img src="https://static.igem.org/mediawiki/2013/2/26/2013ustcigem_IGEM_basic_Logo.png" alt="igem home page" width="50" height="40" /></a><br />
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<div class="content" align="center"> <br />
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<div class="home-pic"><br />
<div class="conbar1"><br />
<div id="breadcrumb"></div><br />
</div><br />
<div class="page-tittle" align="right"><br />
<h1>In Situ <span>T</span>ransdermal <span>Vaccine</span></h1><br />
</div><br />
<div class="page-para" align="right"><br />
<p>Fresh Expression, No Purification<br />Standardized, Block-based Design<br />Easy-transportation, Low Storage External Costs<br />Almost No Demand For Medical Conditions And Proferssionals<br /><br />
</p><p></p><br />
<p><br />
<a class="button-home" href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview"><span>Project </span></a><br />
</p><br />
</div><br />
</div><br />
</div><br />
</div><br />
<br />
<div class="content" id="homecol" align="center"><br />
<div class="conbar2"><br />
<div id="home" class="leftbar" align="left"><br />
<div id="abstract-bar"><br />
<h1>Motivation</h1><br />
<div id="abs-1"><br />
<img class="img-left" src="https://static.igem.org/mediawiki/2013/0/02/2013ustc-china_66.png" width="16" height="14" /><br />
<div id="abs-text"><br />
We hope to develop a fresh-expression transdermal vaccination.<br />
<br />We hope everyone in this world could equally enjoy the benefit of medical progress.<br /><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/Project/Background">more</a><br />
</div><br />
<img class="img-right" src="https://static.igem.org/mediawiki/2013/8/8e/2013ustc-china_99.png" width="16" height="13" /></div><br />
</div><br />
<br />
<br />
<div id="pro-bar"><br />
<h1>Design of Project</h1><br />
<div id="pro-text"><br />
With the support of <span><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Background#" target="_blank">TD-1</a></span>, a special polypeptide which can greatly facilitate macromolecule transdermal delivery through intact skin, we construct three different fuctional engineered bacillus subtilis; besides, we design a reporter system, including a kill switch, in the fouth type B.subtilis to improve usability and ensure biosafety.</div><br />
<div id="ls" class="pro-col"><br />
<img class="img-left" src="https://static.igem.org/mediawiki/2013/0/02/2013ustc-china_66.png" width="16" height="14" /><br />
<div id="pro-de-text"><br />
<span>ANTIGEN</span><br /><br />
As the core of our in-situ system,<br /> TD1-antigen expresser is designed<br /> and will share a great percentage of<br /> the total bacteria in our "band-aid".<br />
<br /><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/Project/Design">more</a><br />
</div><br />
<img class="img-right" src="https://static.igem.org/mediawiki/2013/8/8e/2013ustc-china_99.png" width="16" height="13" /><br />
</div><br />
<div class="pro-col"><br />
<img class="img-left" src="https://static.igem.org/mediawiki/2013/0/02/2013ustc-china_66.png" width="16" height="14" /><br />
<div id="pro-de-text"><br />
<span>LTB</span><br /><br />
As one kind of our adjuvant expresser,<br />TD1-LTB is used to enhance the antigenicity.<br /> For LTB has so many advantages, it's a wild using adjuvant for most of exsiting injected vaccine.<br />
<br /><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/Project/Design">more</a><br />
</div><br />
<img class="img-right" src="https://static.igem.org/mediawiki/2013/8/8e/2013ustc-china_99.png" width="16" height="13" /><br />
</div><br />
<div id="ls" class="pro-col"><br />
<img class="img-left" src="https://static.igem.org/mediawiki/2013/0/02/2013ustc-china_66.png" width="16" height="14" /><br />
<div id="pro-de-text"><br />
<span>TNFα</span><br /><br />
TNFαis a chemotactic factor that recruits Langerhans Cells, antigen presenting cells beneath the skin barrier. The TNFα expresser is responsible for immigrating LC and triggering more intense immune responses.<br />
<br /><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/Project/Design">more</a><br />
</div><br />
<img class="img-right" src="https://static.igem.org/mediawiki/2013/8/8e/2013ustc-china_99.png" width="16" height="13" /><br />
</div><br />
<div class="pro-col"><br />
<img class="img-left" src="https://static.igem.org/mediawiki/2013/0/02/2013ustc-china_66.png" width="16" height="14" /><br />
<div id="pro-de-text"><br />
<span>REPORTER</span><br /><br />
Reporter notifies users whether the status of vaccine patch is all right and when they can stick the patches to arms . Further more killing switch is conveyed in them as a biosafety guard.<br />
<br /><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/Project/Design">more</a><br />
</div><br />
<img class="img-right" src="https://static.igem.org/mediawiki/2013/8/8e/2013ustc-china_99.png" width="16" height="13" /><br />
</div><br />
</div><br />
<br />
<div id="exp-bar"><br />
<h1>Experimental Measurement</h1><br />
<div class="exp-col"><br />
<img class="img-left" src="https://static.igem.org/mediawiki/2013/0/02/2013ustc-china_66.png" width="16" height="14" /><br />
<div id="exp-text"><br />
<span>Molecules part</span><br /><br />
We took E.coli and the PET vector as the positive control and realized our ideas in the B.subtilis expression system based on PHT43 shuttle vector.<br /><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">more</a><br />
</div><br />
<img class="img-right" src="https://static.igem.org/mediawiki/2013/8/8e/2013ustc-china_99.png" width="16" height="13" /><br />
</div><br />
<div class="exp-col"><br />
<img class="img-left" src="https://static.igem.org/mediawiki/2013/0/02/2013ustc-china_66.png" width="16" height="14" /><br />
<div id="exp-text"><br />
<span>Protein part</span><br /><br />
Before we achieve fresh expression, we use purified protein to obtain accurate data to verify our assumptions.<br /><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">more</a><br />
</div><br />
<img class="img-right" src="https://static.igem.org/mediawiki/2013/8/8e/2013ustc-china_99.png" width="16" height="13" /><br />
</div><br />
<div class="exp-col"><br />
<img class="img-left" src="https://static.igem.org/mediawiki/2013/0/02/2013ustc-china_66.png" width="16" height="14" /><br />
<div id="exp-text"><br />
<span>Mice experiments</span><br /><br />
The most obvious character of our 2013USTC iGEM team is that we have not only achieved a series of molecule experiments but also tried mice experiments, which could be divided into transdermal experiments in vitro and animate mice experiments<br /><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results/FurthurWork">more</a><br />
</div><br />
<img class="img-right" src="https://static.igem.org/mediawiki/2013/8/8e/2013ustc-china_99.png" width="16" height="13" /><br />
</div><br />
</div><br />
<br />
<br />
<br />
</div><br />
<div id="home" class="rightbar" style="position:static;" align="left"><br />
<br />
<div id="modle-bar"><br />
<h1>Modeling</h1><br />
<div id="modle-col"><br />
<h2><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Suicide mechanism of the reporter system</a></h2><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch"><img src="https://static.igem.org/mediawiki/2013/4/48/2013ustc-china_Model1.png" width="310" height="160" /></a><br />
<br /><br />
<h2><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">The optimal culture conditions of Bacillus subtilis</a></h2><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture"><img src="https://static.igem.org/mediawiki/2013/1/10/2013ustc-china_Model2.png" width="310" height="234" /></a><br />
<h2><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments"><br />
Designs of Immune Experiments</a></h2><br />
</div><br />
</div><br />
<br />
<div id="hum-bar"><br />
<h1>Human Practice</h1><br />
<div id="hum-col"><br />
<h2><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication"><br />
Communication</a></h2><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication"><img src="https://static.igem.org/mediawiki/2013/5/5e/2013ustc-china_China-map.png" width="310" height="259" /></a><br />
<h2><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication"><br />
Activity</a></h2><br />
</a><br />
</div><br />
</div><br />
<br />
</div><br />
<br />
</div></div><br />
<br />
<br />
<div id="footer"><br />
<div id="thanks"><br />
<h3 id="sponsors-title" align="center"><em>Thanks to the sponsors</em></h3><br><br />
<div class="sponsor" id="db"><br />
<p align="center">Deutsche Bank, China</p><br />
<div align="center"><a href="https://china.db.com/index_e.html" target="_blank"><img src="https://static.igem.org/mediawiki/2013/a/a7/2013ustc-china_Db-logo.png" alt="Deutsche Bank, China" height="65px" align="absmiddle" width="280px"></a><br />
</div><br />
</div><br />
<br />
<div class="sponsor"><br />
<p align="center">School of Life Sciences, USTC</p><br />
<div align="center"><a href="http://en.biox.ustc.edu.cn/" target="_blank"><img src="https://static.igem.org/mediawiki/2013/9/98/2013ustc-china_Life-logo.png" alt="School of Life Sciences, USTC" height="65px" align="absmiddle" width="280px"></a><br />
</div><br />
</div><br />
<br />
<div class="sponsor"><br />
<p align="center">USTC<span style="color:#fcc644">IF</span></p><br />
<div align="center"><a href="http://www.ustcif.org/" target="_blank"><img src="https://static.igem.org/mediawiki/2013/3/3c/2013ustc-china_Ustcif-logo.png" height="65px" align="absmiddle" width="280px"></a><br />
</div><br />
</div><br />
<br />
<div class="sponsor"><br />
<p align="center">School of the Gifted Young, USTC</p><br />
<div align="center"><a href="http://en.scgy.ustc.edu.cn/" target="_blank"><img src="https://static.igem.org/mediawiki/2013/c/c1/2013ustc-china_Gifted_young.jpg" height="65px" align="absmiddle" width="280px"></a><br />
</div><br />
</div><br />
<br />
<div class="sponsor"><br />
<p align="center">School of physics, USTC</p><br />
<div align="center"><a href="http://en.physics.ustc.edu.cn/" target="_blank"><img src="https://static.igem.org/mediawiki/2013/6/68/2013ustc-china_Physics-logo.png" height="65px" align="absmiddle" width="280px"></a><br />
</div><br />
</div><br />
<br />
<div class="sponsor"><br />
<p align="center">Teaching Affairs Office of USTC</p><br />
<div align="center"><a href="http://www.teach.ustc.edu.cn/" target="_blank"><img src="https://static.igem.org/mediawiki/2013/f/f9/2013ustc-china_Teaching-logo.png" alt="Teaching Affairs Office Of USTC" style="background-color:white" height="65px" align="absmiddle" width="280px"></a><br />
</div><br />
</div><br />
</div><br />
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</body><br />
</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/HumanPracticeTeam:USTC CHINA/HumanPractice2013-09-27T22:38:32Z<p>Surine: </p>
<hr />
<div>{{USTC-China/hidden}}<br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li><br />
</ul><br />
</li><br />
<li class="active"><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a><br />
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<div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" /><br />
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<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a></div></div><br />
<div class="part"><br />
<div class="part-tittle"><h>Communication</h></div><br />
<br />
<div class="part-content"><br />
<p>USTC_CHINA 2013<br><br />
Travel almost<span> all </span>over China within half a year<br><br />
Visit<span> Five </span>igem teams,<span> Four </span>CAS institutes,<span> Seventeen </span>senior professors<br><br />
Give special report in<span> Four </span>universities and establish a preliminary cooperative relationship with<span> One </span>famous biotechnology company.<br></p></div><br />
<div class="part-details"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication">Details</a><br />
</div><br />
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<br />
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<img width="804" height="733" style="float:left;" src="https://static.igem.org/mediawiki/2013/5/5e/2013ustc-china_China-map.png" alt="China Map"><br />
<br />
<div class="pin pin-down" data-xpos="575" data-ypos="235"> <br />
<h2><a href="https://2013.igem.org/Team:Peking">PKU iGEM</a> and <a href="http://www.chinacdc.cn/en/aboutus/">CCDCP</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="658" data-ypos="403"> <br />
<h2><a href="https://2013.igem.org/Team:SJTU-BioX-Shanghai">SJTU iGEM</a> and <a href="https://2013.igem.org/Team:Fudan">FDU iGEM</a></h2> <br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="643" data-ypos="376"> <br />
<h2><a href="https://2013.igem.org/Team:NJU_China">NJU iGEM</a> and <a href="http://english.njau.edu.cn/">NJAU</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="602" data-ypos="408"> <br />
<h2><a href="https://2013.igem.org/Team:USTC_CHINA">USTC iGEM</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="544" data-ypos="363"> <br />
<h2><a href="http://www.htu.cn/english/">HNNU</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="530" data-ypos="424"> <br />
<h2><a href="https://2013.igem.org/Team:WHU-China">WHU iGEM</a> , <a href="http://english.whiov.cas.cn/">VRICAS</a> and <a href="http://english.ihb.cas.cn/">CASIAO</a></h2><br />
</div><br />
<div class="pin pin-down" data-xpos="556" data-ypos="568"> <br />
<h2><a href="https://2013.igem.org/Asia">iGEM Asia</a></h2> <br />
</div><br />
<div class="pin pin-down" data-xpos="538" data-ypos="559"> <br />
<h2><a href="http://www.gzpiri.com/eweb/eindex.asp">GPRI</a></h2><br />
</div><br />
</div><br />
<div class="part"><br />
<div class="part-tittle"><h>Activity</h></div><br />
<div class="part-content"><br />
<p>USTC_CHINA 2013</br><br />
Tadditional lab open day,<span> May Festival </span>,receiving more than<span> 500 </span>visitors.<br><br />
Novel campus activity, <span>Super Taste</span>, publicizing synthetic biology to the future elites.</p></div><br />
<div class="img-left-half"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity"><img src="https://static.igem.org/mediawiki/igem.org/1/10/2013ustc-china_may_festival_3.jpg" width="450" height="325"/></a><br />
<div style="width:450px;" align="center">May Festival</div><br />
</div><br />
<div class="img-left-half"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity"><img src="https://static.igem.org/mediawiki/igem.org/e/ea/2013ustc-china_taster_2.jpg"width="450"height="325"/></a><br />
<div style="width:450px;" align="center">Super Taste</div><br />
</div><br />
<div class="part-details"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Details</a><br />
</div><br />
</div> <br />
<br />
<br />
<br />
</div><br />
</body><br />
</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/HumanPracticeTeam:USTC CHINA/HumanPractice2013-09-27T22:35:07Z<p>Surine: </p>
<hr />
<div>{{USTC-China/hidden}}<br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Design">Design</a></li><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li><br />
</ul><br />
</li><br />
<li class="active"><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li><br />
</ul><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Members</a></li><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Attributions">Attributions</a></li><br />
<li><a href="https://igem.org/2013_Judging_Form?id=1074#iGEM_Medals">Achievements</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li><br />
</ul><br />
<div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" /><br />
</div><br />
</div><br />
</div><br />
<br />
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<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a></div></div><br />
<div class="part"><br />
<div class="part-tittle"><h>Communication</h></div><br />
<br />
<div class="part-content"><br />
<p>USTC_CHINA 2013<br><br />
Travel almost<span> all </span>over China within half a year<br><br />
Visit<span> Five </span>igem teams,<span> Four </span>CAS institutes,<span> Seventeen </span>senior professors<br><br />
Give special report in<span> Four </span>universities and establish a preliminary cooperative relationship with<span> One </span>famous biotechnology company.<br></p></div><br />
<div class="part-details"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication">Details</a><br />
</div><br />
</div><br />
<br />
<div id="wrapper"><br />
<img width="804" height="733" style="float:left;" src="https://static.igem.org/mediawiki/2013/5/5e/2013ustc-china_China-map.png" alt="China Map"><br />
<br />
<div class="pin pin-down" data-xpos="575" data-ypos="235"> <br />
<h2><a href="https://2013.igem.org/Team:Peking">PKU iGEM</a> and <a href="http://www.chinacdc.cn/en/aboutus/">CCDCP</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="658" data-ypos="403"> <br />
<h2><a href="https://2013.igem.org/Team:SJTU-BioX-Shanghai">SJTU iGEM</a> and <a href="https://2013.igem.org/Team:Fudan">FDU iGEM</a></h2> <br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="643" data-ypos="376"> <br />
<h2><a href="https://2013.igem.org/Team:NJU_China">NJU iGEM</a> and <a href="http://english.njau.edu.cn/">NJAU</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="602" data-ypos="408"> <br />
<h2><a href="https://2013.igem.org/Team:USTC_CHINA">USTC iGEM</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="544" data-ypos="363"> <br />
<h2><a href="http://www.htu.cn/english/">HNNU</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="530" data-ypos="424"> <br />
<h2><a href="https://2013.igem.org/Team:WHU-China">WHU iGEM</a> , <a href="http://english.whiov.cas.cn/">VRICAS</a> and <a href="http://english.ihb.cas.cn/">CASIAO</a></h2><br />
</div><br />
<div class="pin pin-down" data-xpos="556" data-ypos="568"> <br />
<h2><a href="https://2013.igem.org/Asia">iGEM Asia</a></h2> <br />
</div><br />
<div class="pin pin-down" data-xpos="538" data-ypos="559"> <br />
<h2><a href="http://www.gzpiri.com/eweb/eindex.asp">GPRI</a></h2><br />
</div><br />
</div><br />
<div class="part"><br />
<div class="part-tittle"><h>Activity</h></div><br />
<div class="part-content"><br />
<p>USTC_CHINA 2013</br><br />
Tadditional lab open day,<span> May Festival </span>,receiving more than<span> 500 </span>visitors.<br><br />
Novel campus activity, <span>Super Taste</span>, publicizing synthetic biology to the future elites.</p></div><br />
<div class="img-left-half"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity"><img src="https://static.igem.org/mediawiki/igem.org/1/10/2013ustc-china_may_festival_3.jpg" width="450" height="325"/></a><br />
<div style="width:450px;" align="center">May Festival</div><br />
</div><br />
<div class="img-left-half"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity"><img src="https://static.igem.org/mediawiki/igem.org/e/ea/2013ustc-china_taster_2.jpg"width="450"height="325"/></a><br />
<div style="width:450px;" align="center">Super Taste/div><br />
</div><br />
<div class="part-details"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Details</a><br />
</div><br />
</div> <br />
<br />
<br />
<br />
</div><br />
</body><br />
</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/humanpractice.cssTeam:USTC CHINA/humanpractice.css2013-09-27T22:35:02Z<p>Surine: </p>
<hr />
<div>@charset "utf-8";<br />
/* CSS Document */<br />
.part{<br />
width:964px;<br />
height:185px;<br />
text-align:left;<br />
position:relative;<br />
}<br />
.part-tittle{<br />
width:964px;<br />
height:16px;<br />
padding-bottom:4px;<br />
}<br />
.content .part .part-tittle h{<br />
float:left;<br />
font-size:24px;<br />
text-indent:3px;<br />
color:#303030;<br />
}<br />
.part-content{<br />
width:964px;<br />
}<br />
.part-details{<br />
width:24px;<br />
}<br />
<br />
.part-details a{<br />
padding-right:6px;<br />
padding-bottom:4px;<br />
font-size:16px;<br />
text-indent:3px;<br />
float:left;<br />
position:relative;<br />
z-index:10;<br />
}<br />
<br />
<br />
.content .part-content p{<br />
float:left;<br />
font-size:16px;<br />
margin-left:3px;<br />
line-height:25px;<br />
color:#797979;<br />
}<br />
.content .part-content p span{<br />
font-size:20px;<br />
color:#ff9933;<br />
}<br />
.img-left-half{<br />
float:left;width:450px;padding15px;<br />
}</div>Surinehttp://2013.igem.org/Team:USTC_CHINA/HumanPracticeTeam:USTC CHINA/HumanPractice2013-09-27T22:24:30Z<p>Surine: </p>
<hr />
<div>{{USTC-China/hidden}}<br />
<html><br />
<head><br />
<link rel="stylesheet" type="text/css" href="https://2013.igem.org/Team:USTC_CHINA/main.css?action=raw&ctype=text/css" /><br />
<link rel="stylesheet" type="text/css" href="https://2013.igem.org/Team:USTC_CHINA/humanpractice.css?action=raw&ctype=text/css" /><br />
<script src="http://code.jquery.com/jquery-1.6.3.min.js" type= "text/javascript"> </script><br />
<script><br />
$(document).ready(function(){<br />
<br />
// set the wrapper width and height to match the img size<br />
$('#wrapper').css({'width':$('#wrapper img').width(),<br />
'height':$('#wrapper img').height()<br />
})<br />
<br />
//tooltip direction<br />
var tooltipDirection;<br />
<br />
for (i=0; i<$(".pin").length; i++)<br />
{ <br />
// set tooltip direction type - up or down <br />
if ($(".pin").eq(i).hasClass('pin-down')) {<br />
tooltipDirection = 'tooltip-down';<br />
} else {<br />
tooltipDirection = 'tooltip-up';<br />
}<br />
<br />
// append the tooltip<br />
$("#wrapper").append("<div style='left:"+$(".pin").eq(i).data('xpos')+"px;top:"+$(".pin").eq(i).data('ypos')+"px' class='" + tooltipDirection +"'>\<br />
<div class='tooltip'>" + $(".pin").eq(i).html() + "</div>\<br />
</div>");<br />
} <br />
<br />
// show/hide the tooltip<br />
$('.tooltip-up, .tooltip-down').mouseenter(function(){<br />
$(this).children('.tooltip').fadeIn(100);<br />
}).mouseleave(function(){<br />
$(this).children('.tooltip').fadeOut(100);<br />
})<br />
});<br />
</script><br />
<style><br />
body {<br />
text-align: center;<br />
font: 13px Arial,Helvetica; <br />
}<br />
<br />
/* Relative positioning*/<br />
#wrapper {<br />
position: relative;<br />
margin: 0px 0px 20px -146px;<br />
border: 1px solid #fafafa;<br />
-moz-box-shadow: 0 3px 3px rgba(0,0,0,.5);<br />
-webkit-box-shadow: 0 3px 3px rgba(0,0,0,.5);<br />
box-shadow: 0 3px 3px rgba(0,0,0,.5);<br />
} <br />
/* Hide the original tooltips contents */<br />
.pin {<br />
display: none;<br />
} <br />
/* Begin styling the tooltips and pins */<br />
.tooltip-up, .tooltip-down {<br />
position: absolute;<br />
background: url("https://static.igem.org/mediawiki/2013/9/97/T-pointer.png");<br />
width: 40px;<br />
height: 52px;<br />
} <br />
<br />
<br />
.tooltip {<br />
display: none;<br />
width: 200px;<br />
cursor: help;<br />
text-shadow: 0 1px 0 #fff;<br />
position: absolute;<br />
top: 10px;<br />
left: 50%;<br />
z-index: 999;<br />
margin-left: -115px;<br />
padding:15px;<br />
color: #222;<br />
-moz-border-radius: 5px;<br />
-webkit-border-radius: 5px;<br />
border-radius: 5px;<br />
-moz-box-shadow: 0 3px 0 rgba(0,0,0,.7);<br />
-webkit-box-shadow: 0 3px 0 rgba(0,0,0,.7);<br />
box-shadow: 0 3px 0 rgba(0,0,0,.7);<br />
background: #fff1d3;<br />
background: -webkit-gradient(linear, left top, left bottom, from(#fff1d3), to(#ffdb90));<br />
background: -webkit-linear-gradient(top, #fff1d3, #ffdb90);<br />
background: -moz-linear-gradient(top, #fff1d3, #ffdb90);<br />
background: -ms-linear-gradient(top, #fff1d3, #ffdb90);<br />
background: -o-linear-gradient(top, #fff1d3, #ffdb90);<br />
background: linear-gradient(top, #fff1d3, #ffdb90); <br />
}<br />
<br />
.tooltip::after {<br />
content: '';<br />
position: absolute;<br />
top: -10px;<br />
left: 50%;<br />
margin-left: -10px;<br />
border-bottom: 10px solid #fff1d3;<br />
border-left: 10px solid transparent;<br />
border-right :10px solid transparent;<br />
}<br />
.tooltip-down .tooltip {<br />
bottom: 12px;<br />
top: auto;<br />
}<br />
<br />
.tooltip-down .tooltip::after {<br />
bottom: -10px;<br />
top: auto;<br />
border-bottom: 0;<br />
border-top: 10px solid #ffdb90;<br />
}<br />
<br />
.tooltip h2 {<br />
font: bold 1.3em 'Trebuchet MS', Tahoma, Arial;<br />
margin: 0 0 10px;<br />
}<br />
<br />
.tooltip ul {<br />
margin: 0;<br />
padding: 0;<br />
list-style: none;<br />
} <br />
</style><br />
</head><br />
<body background="https://static.igem.org/mediawiki/2013/6/62/2013ustc-china_Light_grey_bg.png"><br />
<div class="bar" align="center"><br />
<div class="container" align="left"><br />
<div id="igemlogo"><a href="https://2013.igem.org/Main_Page" target="_blank"><img src="https://static.igem.org/mediawiki/2013/2/26/2013ustcigem_IGEM_basic_Logo.png" alt="igem home page" width="50" height="40" /></a></div><br />
<br />
<ul id="nav"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Background">Background</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Design">Design</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">Notebook</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">Timeline</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li><br />
</ul><br />
</li><br />
<li class="active"><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Members</a></li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=USTC_CHINA">Profile</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Attributions">Attributions</a></li><br />
<li><a href="https://igem.org/2013_Judging_Form?id=1074#iGEM_Medals">Achievements</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li><br />
</ul><br />
<div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" /><br />
</div><br />
</div><br />
</div><br />
<br />
<div class="content" align="center"><br />
<div class="conbar1"><br />
<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a></div></div><br />
<div class="part"><br />
<div class="part-tittle"><h>Communication</h></div><br />
<br />
<div class="part-content"><br />
<p>USTC_CHINA 2013<br><br />
Travel almost<span> all </span>over China within half a year<br><br />
Visit<span> Five </span>igem teams,<span> Four </span>CAS institutes,<span> Seventeen </span>senior professors<br><br />
Give special report in<span> Four </span>universities and establish a preliminary cooperative relationship with<span> One </span>famous biotechnology company.<br></p></div><br />
<div class="part-details"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication">Details</a><br />
</div><br />
</div><br />
<br />
<div id="wrapper"><br />
<img width="804" height="733" style="float:left;" src="https://static.igem.org/mediawiki/2013/5/5e/2013ustc-china_China-map.png" alt="China Map"><br />
<br />
<div class="pin pin-down" data-xpos="575" data-ypos="235"> <br />
<h2><a href="https://2013.igem.org/Team:Peking">PKU iGEM</a> and <a href="http://www.chinacdc.cn/en/aboutus/">CCDCP</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="658" data-ypos="403"> <br />
<h2><a href="https://2013.igem.org/Team:SJTU-BioX-Shanghai">SJTU iGEM</a> and <a href="https://2013.igem.org/Team:Fudan">FDU iGEM</a></h2> <br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="643" data-ypos="376"> <br />
<h2><a href="https://2013.igem.org/Team:NJU_China">NJU iGEM</a> and <a href="http://english.njau.edu.cn/">NJAU</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="602" data-ypos="408"> <br />
<h2><a href="https://2013.igem.org/Team:USTC_CHINA">USTC iGEM</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="544" data-ypos="363"> <br />
<h2><a href="http://www.htu.cn/english/">HNNU</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="530" data-ypos="424"> <br />
<h2><a href="https://2013.igem.org/Team:WHU-China">WHU iGEM</a> , <a href="http://english.whiov.cas.cn/">VRICAS</a> and <a href="http://english.ihb.cas.cn/">CASIAO</a></h2><br />
</div><br />
<div class="pin pin-down" data-xpos="556" data-ypos="568"> <br />
<h2><a href="https://2013.igem.org/Asia">iGEM Asia</a></h2> <br />
</div><br />
<div class="pin pin-down" data-xpos="538" data-ypos="559"> <br />
<h2><a href="http://www.gzpiri.com/eweb/eindex.asp">GPRI</a></h2><br />
</div><br />
</div><br />
<div class="part"><br />
<div class="part-tittle"><h>Activity</h></div><br />
<div class="part-content"><br />
<p>USTC_CHINA 2013</br><br />
Tadditional lab open day,<span> May Festival </span>,receiving more than<span> 500 </span>visitors.<br><br />
Novel campus activity, <span>Super Taste</span>, publicizing synthetic biology to the future elites.</p></div><br />
<div class="part-details"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Details</a><br />
</div><br />
</div> <br />
<br />
<br />
<br />
</div><br />
</body><br />
</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/HumanPracticeTeam:USTC CHINA/HumanPractice2013-09-27T22:24:07Z<p>Surine: </p>
<hr />
<div>{{USTC-China/hidden}}<br />
<html><br />
<head><br />
<link rel="stylesheet" type="text/css" href="https://2013.igem.org/Team:USTC_CHINA/main.css?action=raw&ctype=text/css" /><br />
<link rel="stylesheet" type="text/css" href="https://2013.igem.org/Team:USTC_CHINA/humanpractice.css?action=raw&ctype=text/css" /><br />
<script src="http://code.jquery.com/jquery-1.6.3.min.js" type= "text/javascript"> </script><br />
<script><br />
$(document).ready(function(){<br />
<br />
// set the wrapper width and height to match the img size<br />
$('#wrapper').css({'width':$('#wrapper img').width(),<br />
'height':$('#wrapper img').height()<br />
})<br />
<br />
//tooltip direction<br />
var tooltipDirection;<br />
<br />
for (i=0; i<$(".pin").length; i++)<br />
{ <br />
// set tooltip direction type - up or down <br />
if ($(".pin").eq(i).hasClass('pin-down')) {<br />
tooltipDirection = 'tooltip-down';<br />
} else {<br />
tooltipDirection = 'tooltip-up';<br />
}<br />
<br />
// append the tooltip<br />
$("#wrapper").append("<div style='left:"+$(".pin").eq(i).data('xpos')+"px;top:"+$(".pin").eq(i).data('ypos')+"px' class='" + tooltipDirection +"'>\<br />
<div class='tooltip'>" + $(".pin").eq(i).html() + "</div>\<br />
</div>");<br />
} <br />
<br />
// show/hide the tooltip<br />
$('.tooltip-up, .tooltip-down').mouseenter(function(){<br />
$(this).children('.tooltip').fadeIn(100);<br />
}).mouseleave(function(){<br />
$(this).children('.tooltip').fadeOut(100);<br />
})<br />
});<br />
</script><br />
<style><br />
body {<br />
text-align: center;<br />
font: 13px Arial,Helvetica; <br />
}<br />
<br />
/* Relative positioning*/<br />
#wrapper {<br />
position: relative;<br />
margin: 0px 0px 20px -146px;<br />
border: 1px solid #fafafa;<br />
-moz-box-shadow: 0 3px 3px rgba(0,0,0,.5);<br />
-webkit-box-shadow: 0 3px 3px rgba(0,0,0,.5);<br />
box-shadow: 0 3px 3px rgba(0,0,0,.5);<br />
} <br />
/* Hide the original tooltips contents */<br />
.pin {<br />
display: none;<br />
} <br />
/* Begin styling the tooltips and pins */<br />
.tooltip-up, .tooltip-down {<br />
position: absolute;<br />
background: url("https://static.igem.org/mediawiki/2013/9/97/T-pointer.png");<br />
width: 40px;<br />
height: 52px;<br />
} <br />
<br />
<br />
.tooltip {<br />
display: none;<br />
width: 200px;<br />
cursor: help;<br />
text-shadow: 0 1px 0 #fff;<br />
position: absolute;<br />
top: 10px;<br />
left: 50%;<br />
z-index: 999;<br />
margin-left: -115px;<br />
padding:15px;<br />
color: #222;<br />
-moz-border-radius: 5px;<br />
-webkit-border-radius: 5px;<br />
border-radius: 5px;<br />
-moz-box-shadow: 0 3px 0 rgba(0,0,0,.7);<br />
-webkit-box-shadow: 0 3px 0 rgba(0,0,0,.7);<br />
box-shadow: 0 3px 0 rgba(0,0,0,.7);<br />
background: #fff1d3;<br />
background: -webkit-gradient(linear, left top, left bottom, from(#fff1d3), to(#ffdb90));<br />
background: -webkit-linear-gradient(top, #fff1d3, #ffdb90);<br />
background: -moz-linear-gradient(top, #fff1d3, #ffdb90);<br />
background: -ms-linear-gradient(top, #fff1d3, #ffdb90);<br />
background: -o-linear-gradient(top, #fff1d3, #ffdb90);<br />
background: linear-gradient(top, #fff1d3, #ffdb90); <br />
}<br />
<br />
.tooltip::after {<br />
content: '';<br />
position: absolute;<br />
top: -10px;<br />
left: 50%;<br />
margin-left: -10px;<br />
border-bottom: 10px solid #fff1d3;<br />
border-left: 10px solid transparent;<br />
border-right :10px solid transparent;<br />
}<br />
.tooltip-down .tooltip {<br />
bottom: 12px;<br />
top: auto;<br />
}<br />
<br />
.tooltip-down .tooltip::after {<br />
bottom: -10px;<br />
top: auto;<br />
border-bottom: 0;<br />
border-top: 10px solid #ffdb90;<br />
}<br />
<br />
.tooltip h2 {<br />
font: bold 1.3em 'Trebuchet MS', Tahoma, Arial;<br />
margin: 0 0 10px;<br />
}<br />
<br />
.tooltip ul {<br />
margin: 0;<br />
padding: 0;<br />
list-style: none;<br />
} <br />
</style><br />
</head><br />
<body background="https://static.igem.org/mediawiki/2013/6/62/2013ustc-china_Light_grey_bg.png"><br />
<div class="bar" align="center"><br />
<div class="container" align="left"><br />
<div id="igemlogo"><a href="https://2013.igem.org/Main_Page" target="_blank"><img src="https://static.igem.org/mediawiki/2013/2/26/2013ustcigem_IGEM_basic_Logo.png" alt="igem home page" width="50" height="40" /></a></div><br />
<br />
<ul id="nav"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Background">Background</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Design">Design</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">Notebook</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">Timeline</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li><br />
</ul><br />
</li><br />
<li class="active"><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Members</a></li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=USTC_CHINA">Profile</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Attributions">Attributions</a></li><br />
<li><a href="https://igem.org/2013_Judging_Form?id=1074#iGEM_Medals">Achievements</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li><br />
</ul><br />
<div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" /><br />
</div><br />
</div><br />
</div><br />
<br />
<div class="content" align="center"><br />
<div class="conbar1"><br />
<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a></div></div><br />
<div class="part"><br />
<div class="part-tittle"><h>Communication</h></div><br />
<br />
<div class="part-content"><br />
<p>USTC_CHINA 2013<br><br />
Travel almost<span> all </span>over China within half a year<br><br />
Visit<span> Five </span>igem teams,<span> Four </span>CAS institutes,<span> Seventeen </span>senior professors<br><br />
Give special report in<span> Four </span>universities and establish a preliminary cooperative relationship with<span> One </span>famous biotechnology company.<br></p></div><br />
<div class="part-details"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication">Details</a><br />
</div><br />
</div><br />
<br />
<div id="wrapper"><br />
<img width="804" height="733" style="float:left;" src="https://static.igem.org/mediawiki/2013/5/5e/2013ustc-china_China-map.png" alt="China Map"><br />
<br />
<div class="pin pin-down" data-xpos="575" data-ypos="235"> <br />
<h2><a href="https://2013.igem.org/Team:Peking">PKU iGEM</a> and <a href="http://www.chinacdc.cn/en/aboutus/">CCDCP</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="658" data-ypos="403"> <br />
<h2><a href="https://2013.igem.org/Team:SJTU-BioX-Shanghai">SJTU iGEM</a> and <a href="https://2013.igem.org/Team:Fudan">FDU iGEM</a></h2> <br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="643" data-ypos="376"> <br />
<h2><a href="https://2013.igem.org/Team:NJU_China">NJU iGEM</a> and <a href="http://english.njau.edu.cn/">NJAU</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="602" data-ypos="408"> <br />
<h2><a href="https://2013.igem.org/Team:USTC_CHINA">USTC iGEM</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="544" data-ypos="363"> <br />
<h2><a href="http://www.htu.cn/english/">HNNU</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="530" data-ypos="424"> <br />
<h2><a href="https://2013.igem.org/Team:WHU-China">WHU iGEM</a> , <a href="http://english.whiov.cas.cn/">VRICAS</a> and <a href="http://english.ihb.cas.cn/">CASIAO</a></h2><br />
</div><br />
<div class="pin pin-down" data-xpos="556" data-ypos="568"> <br />
<h2><a href="https://2013.igem.org/Asia">iGEM Asia</a></h2> <br />
</div><br />
<div class="pin pin-down" data-xpos="538" data-ypos="559"> <br />
<h2><a href="http://www.gzpiri.com/eweb/eindex.asp">GPRI</a></h2><br />
</div><br />
</div><br />
<div class="part"><br />
<div class="part-tittle"><h>Activity</h></div><br />
<div class="part-content"><br />
<p>USTC_CHINA 2013</br><br />
Tadditional lab open day,<span> May Festival </span>,receiving more than<span> 500 </span>visitors.<br />
Novel campus activity, <span>Super Taste</span>, publicizing synthetic biology to the future elites.</p></div><br />
<div class="part-details"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Details</a><br />
</div><br />
</div> <br />
<br />
<br />
<br />
</div><br />
</body><br />
</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/HumanPracticeTeam:USTC CHINA/HumanPractice2013-09-27T22:23:19Z<p>Surine: </p>
<hr />
<div>{{USTC-China/hidden}}<br />
<html><br />
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<link rel="stylesheet" type="text/css" href="https://2013.igem.org/Team:USTC_CHINA/main.css?action=raw&ctype=text/css" /><br />
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<body background="https://static.igem.org/mediawiki/2013/6/62/2013ustc-china_Light_grey_bg.png"><br />
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<div class="container" align="left"><br />
<div id="igemlogo"><a href="https://2013.igem.org/Main_Page" target="_blank"><img src="https://static.igem.org/mediawiki/2013/2/26/2013ustcigem_IGEM_basic_Logo.png" alt="igem home page" width="50" height="40" /></a></div><br />
<br />
<ul id="nav"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Background">Background</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Design">Design</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">Notebook</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">Timeline</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li><br />
</ul><br />
</li><br />
<li class="active"><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Members</a></li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=USTC_CHINA">Profile</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Attributions">Attributions</a></li><br />
<li><a href="https://igem.org/2013_Judging_Form?id=1074#iGEM_Medals">Achievements</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li><br />
</ul><br />
<div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" /><br />
</div><br />
</div><br />
</div><br />
<br />
<div class="content" align="center"><br />
<div class="conbar1"><br />
<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a></div></div><br />
<div class="part"><br />
<div class="part-tittle"><h>Communication</h></div><br />
<br />
<div class="part-content"><br />
<p>USTC_CHINA 2013<br><br />
Travel almost<span> all </span>over China within half a year<br><br />
Visit<span> Five </span>igem teams,<span> Four </span>CAS institutes,<span> Seventeen </span>senior professors<br><br />
Give special report in<span> Four </span>universities and establish a preliminary cooperative relationship with<span> One </span>famous biotechnology company.<br></p></div><br />
<div class="part-details"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication">Details</a><br />
</div><br />
</div><br />
<br />
<div id="wrapper"><br />
<img width="804" height="733" style="float:left;" src="https://static.igem.org/mediawiki/2013/5/5e/2013ustc-china_China-map.png" alt="China Map"><br />
<br />
<div class="pin pin-down" data-xpos="575" data-ypos="235"> <br />
<h2><a href="https://2013.igem.org/Team:Peking">PKU iGEM</a> and <a href="http://www.chinacdc.cn/en/aboutus/">CCDCP</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="658" data-ypos="403"> <br />
<h2><a href="https://2013.igem.org/Team:SJTU-BioX-Shanghai">SJTU iGEM</a> and <a href="https://2013.igem.org/Team:Fudan">FDU iGEM</a></h2> <br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="643" data-ypos="376"> <br />
<h2><a href="https://2013.igem.org/Team:NJU_China">NJU iGEM</a> and <a href="http://english.njau.edu.cn/">NJAU</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="602" data-ypos="408"> <br />
<h2><a href="https://2013.igem.org/Team:USTC_CHINA">USTC iGEM</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="544" data-ypos="363"> <br />
<h2><a href="http://www.htu.cn/english/">HNNU</a></h2><br />
</div><br />
<br />
<div class="pin pin-down" data-xpos="530" data-ypos="424"> <br />
<h2><a href="https://2013.igem.org/Team:WHU-China">WHU iGEM</a> , <a href="http://english.whiov.cas.cn/">VRICAS</a> and <a href="http://english.ihb.cas.cn/">CASIAO</a></h2><br />
</div><br />
<div class="pin pin-down" data-xpos="556" data-ypos="568"> <br />
<h2><a href="https://2013.igem.org/Asia">iGEM Asia</a></h2> <br />
</div><br />
<div class="pin pin-down" data-xpos="538" data-ypos="559"> <br />
<h2><a href="http://www.gzpiri.com/eweb/eindex.asp">GPRI</a></h2><br />
</div><br />
</div><br />
<div class="part"><br />
<div class="part-tittle"><h>Activity</h></div><br />
<div class="part-content"><br />
<p>USTC_CHINA 2013</br><br />
Tadditional lab open day,<span> May Festival <span>,receiving more than<span> 500 </span>visitors.<br />
Novel campus activity, <span>Super Taste<span>, publicizing synthetic biology to the future elites.</p></div><br />
<div class="part-details"><br />
<a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Details</a><br />
</div><br />
</div> <br />
<br />
<br />
<br />
</div><br />
</body><br />
</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/humanpractice.cssTeam:USTC CHINA/humanpractice.css2013-09-27T22:23:13Z<p>Surine: </p>
<hr />
<div>@charset "utf-8";<br />
/* CSS Document */<br />
.part{<br />
width:964px;<br />
height:185px;<br />
text-align:left;<br />
position:relative;<br />
}<br />
.part-tittle{<br />
width:964px;<br />
height:16px;<br />
padding-bottom:4px;<br />
}<br />
.content .part .part-tittle h{<br />
float:left;<br />
font-size:24px;<br />
text-indent:3px;<br />
color:#303030;<br />
}<br />
.part-content{<br />
width:964px;<br />
}<br />
.part-details{<br />
width:24px;<br />
}<br />
<br />
.part-details a{<br />
padding-right:6px;<br />
padding-bottom:4px;<br />
font-size:16px;<br />
text-indent:3px;<br />
float:left;<br />
position:relative;<br />
z-index:10;<br />
}<br />
<br />
<br />
.content .part-content p{<br />
float:left;<br />
font-size:16px;<br />
margin-left:3px;<br />
line-height:25px;<br />
color:#797979;<br />
}<br />
.content .part-content p span{<br />
font-size:20px;<br />
color:#ff9933;<br />
}</div>Surinehttp://2013.igem.org/Team:USTC_CHINA/Project/DesignTeam:USTC CHINA/Project/Design2013-09-27T19:21:18Z<p>Surine: </p>
<hr />
<div>{{USTC-China/hidden}}<br />
<html><br />
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<link rel="stylesheet" type="text/css" href="https://2013.igem.org/Team:USTC_CHINA/main.css?action=raw&ctype=text/css" /><br />
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</head><br />
<body background="https://static.igem.org/mediawiki/2013/6/62/2013ustc-china_Light_grey_bg.png"><br />
<div class="bar" align="center"><br />
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<br />
<div id="igemlogo"><a href="https://2013.igem.org/Main_Page" target="_blank"><img src="https://static.igem.org/mediawiki/2013/2/26/2013ustcigem_IGEM_basic_Logo.png" alt="igem home page" width="50" height="40" /></a></div><br />
<br />
<ul id="nav"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a></li><br />
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<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Background">Background</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Design">Design</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></li><br />
</ul><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">Notebook</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">Timeline</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
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<h1 style="font-size:40px;">Gene Circuit</h1><br />
<div class="basic-bar"><br />
<h2>Introduction</h2><br />
<p>Our T-vaccine consists four modules. Each of them carries a gene circuit independently. The first one expresses TD1-Antigen fusion protein, as the core of our vaccine. The second one expresses TD1-Anjuvant fusion proteins, which enhance the antigenicity. The third one is reporter, it notifies users whether the status of vaccine patch is all right and when they can stick the patches to arms. The last module is a kill switch,we designed a reliable suicide system in bacillus subtilis for the very first time in iGEM. So it would be OK if our users tear patches off and throw them away. We attribute different gene circuits to different bacteria(Modularization idea), by changing the ratio of each bacteria we could make it compatible with any vaccine.</p><br />
<img src="https://static.igem.org/mediawiki/2013/0/0a/2013ustc-chinaWorkflow.png" width="580" height="300" style="margin-left:-30px;"/><br />
<div class="atfirgure" align="center" style="width:580px;">Work Flow</a></div><br />
</div><br />
<div class="basic-bar"><br />
<h2>Our Design</h2> <br />
<img src="https://static.igem.org/mediawiki/2013/4/4d/2013ustc-china_genecircuit.png" width="580" height="180"/><br />
<h3>1. Antigen</h3><br />
<p>As the main part of T-vaccine, TD-1 is fused with different kinds of antigens, so it can penetrate the skin and provoke immune responses. We have designed three kinds of vaccine, which are against hepatitis B, tuberculosis and anthrax. Our ELISA test had proved their antigenicity and mice test will figure it out if they bave immunogenicity,thereby proving that T-vaccine can be generally used. </p><br />
<br><br><br />
<h3>2. Adjuvant</h3><br />
<p>Similar with traditional injection vaccine, adjuvant should be added into vaccine to ensure immunity. We fused TD-1 with LTB. LT has adjuvant activity and can assist foreign antigen to induce the body to produce systemic immune response. The B submit of LTB protein has no toxic effect and it also works.</p><br />
<br><br><br />
<h3>3. TNFα</h3><br />
<p>TNFα can recruit Langerhans cell(LC) which work as antigen-presenting cells around epidermis, and improved LC could transmit into adjacent lymph node provoking the immune response. Because this circuit is quite similar with 1# and 2# ,while its effect might be difficult to certified, so we delayed the schedule of this part. 3# is the only circuit which has not been built or tested during the whole summer experiment.<br />
</div><br />
<br />
<br />
<br />
<div class="basic-bar"><br />
<h2>4. Reporter System</h2><br />
<p>To be more user-friendly, 4# contains a reporting system. After melting in water, the spores will germinate and express blue pigment protein (amilCP) to report the best using time. Meanwhile, 4# could also ensure biosafety, as 4# engineering bacteria can kill all the engineered bacteria after use.</p><br />
<h3>4.1 Reporter</h3><br />
<img src="https://static.igem.org/mediawiki/igem.org/a/aa/2013ustc-china_design111reporter.png" width="580" height="160"/><br />
<p>T-vaccine is also a user-friendly product. Our potential users are not medical professionals, they activate T-vaccine by exposing it to water, and the patch gives visualized signal to notify users whether the status of engineered bacteria is all right and when to stick the patch to arms. We achieve this simply with the regulation of promoter 43, which is recognized by sigma factor A. The activity of promoter 43 is maximal during the exponential growth phase. In other words, we expect engineering Bacillus subtilis to express blue pigment a few hours before expressing massive antigens and adjuvants. It is the perfect time.</p><br />
<br><br><br />
<h3>4.2 Kill Switch</h3><br />
<img src="https://static.igem.org/mediawiki/2013/f/f0/2013ustc-china_sdpABC.png" width="580" height="450"/><br />
<br><br><br />
<p>In B.subtilis, when it comes to the stationary phase, the environmental pressure increases and nutrition becomes limited, so B.subtilis begins to produce spore. Now the community will be divided into two different parts. One of them are trying to kill others to get enough nutrient, delaying the production of spores and achieving a competitive advantage. Killing is mediated by the exported toxic protein SdpC. SdpI will appear on the membrane surface to avoid itself from being damaged. SdpI could bind free SdpC and autopressor SdpR, to remove inhibition of SdpR against I and R, to produce more SdpI to offset SdpC, finally guaranteeing the subgroup alive, thereby delaying the spores production.We transfer SdpC which is fused by promoter SdpI/R into high copy plasmids in order to damage the balance of the system, thereby killing whole colony. When SdpC appears, SdpI on the membrane will bind free SdpC and adsorb SdpR to cease its inhibition against SdpI P/R, trying to produce more SdpI. At the same time, it will activate the promoter SdpR/I in our circuits and generate more SdpC. The system would fall into an infinite loop, and according to our modeling,the amount of SdpC increases beyond the ability of SdpI. Thus, the cells with protection mechanism will crack and die because of too much SdpC. All of them formed the killing device.</p><br />
<img src="https://static.igem.org/mediawiki/igem.org/5/5d/2013ustc-china_design11sdpABC.png" width="580" height="160"/><br />
<p>We Also designed a test circuit, which contains promotor grac and sdpABC only, aiming to determine the ability of SdpC.</p><br />
<br />
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</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/Project/DesignTeam:USTC CHINA/Project/Design2013-09-27T19:20:44Z<p>Surine: </p>
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<h1 style="font-size:40px;">Gene Circuit</h1><br />
<div class="basic-bar"><br />
<h2>Introduction</h2><br />
<p>Our T-vaccine consists four modules. Each of them carries a gene circuit independently. The first one expresses TD1-Antigen fusion protein, as the core of our vaccine. The second one expresses TD1-Anjuvant fusion proteins, which enhance the antigenicity. The third one is reporter, it notifies users whether the status of vaccine patch is all right and when they can stick the patches to arms. The last module is a kill switch,we designed a reliable suicide system in bacillus subtilis for the very first time in iGEM. So it would be OK if our users tear patches off and throw them away. We attribute different gene circuits to different bacteria(Modularization idea), by changing the ratio of each bacteria we could make it compatible with any vaccine.</p><br />
<img src="https://static.igem.org/mediawiki/2013/0/0a/2013ustc-chinaWorkflow.png" width="580" height="300" style="margin-left:-30px;"/><br />
<div class="atfirgure" align="center" style="width:580px;">Work Flow</a><br />
</div><br />
<div class="basic-bar"><br />
<h2>Our Design</h2> <br />
<img src="https://static.igem.org/mediawiki/2013/4/4d/2013ustc-china_genecircuit.png" width="580" height="180"/><br />
<h3>1. Antigen</h3><br />
<p>As the main part of T-vaccine, TD-1 is fused with different kinds of antigens, so it can penetrate the skin and provoke immune responses. We have designed three kinds of vaccine, which are against hepatitis B, tuberculosis and anthrax. Our ELISA test had proved their antigenicity and mice test will figure it out if they bave immunogenicity,thereby proving that T-vaccine can be generally used. </p><br />
<br><br><br />
<h3>2. Adjuvant</h3><br />
<p>Similar with traditional injection vaccine, adjuvant should be added into vaccine to ensure immunity. We fused TD-1 with LTB. LT has adjuvant activity and can assist foreign antigen to induce the body to produce systemic immune response. The B submit of LTB protein has no toxic effect and it also works.</p><br />
<br><br><br />
<h3>3. TNFα</h3><br />
<p>TNFα can recruit Langerhans cell(LC) which work as antigen-presenting cells around epidermis, and improved LC could transmit into adjacent lymph node provoking the immune response. Because this circuit is quite similar with 1# and 2# ,while its effect might be difficult to certified, so we delayed the schedule of this part. 3# is the only circuit which has not been built or tested during the whole summer experiment.<br />
</div><br />
<br />
<br />
<br />
<div class="basic-bar"><br />
<h2>4. Reporter System</h2><br />
<p>To be more user-friendly, 4# contains a reporting system. After melting in water, the spores will germinate and express blue pigment protein (amilCP) to report the best using time. Meanwhile, 4# could also ensure biosafety, as 4# engineering bacteria can kill all the engineered bacteria after use.</p><br />
<h3>4.1 Reporter</h3><br />
<img src="https://static.igem.org/mediawiki/igem.org/a/aa/2013ustc-china_design111reporter.png" width="580" height="160"/><br />
<p>T-vaccine is also a user-friendly product. Our potential users are not medical professionals, they activate T-vaccine by exposing it to water, and the patch gives visualized signal to notify users whether the status of engineered bacteria is all right and when to stick the patch to arms. We achieve this simply with the regulation of promoter 43, which is recognized by sigma factor A. The activity of promoter 43 is maximal during the exponential growth phase. In other words, we expect engineering Bacillus subtilis to express blue pigment a few hours before expressing massive antigens and adjuvants. It is the perfect time.</p><br />
<br><br><br />
<h3>4.2 Kill Switch</h3><br />
<img src="https://static.igem.org/mediawiki/2013/f/f0/2013ustc-china_sdpABC.png" width="580" height="450"/><br />
<br><br><br />
<p>In B.subtilis, when it comes to the stationary phase, the environmental pressure increases and nutrition becomes limited, so B.subtilis begins to produce spore. Now the community will be divided into two different parts. One of them are trying to kill others to get enough nutrient, delaying the production of spores and achieving a competitive advantage. Killing is mediated by the exported toxic protein SdpC. SdpI will appear on the membrane surface to avoid itself from being damaged. SdpI could bind free SdpC and autopressor SdpR, to remove inhibition of SdpR against I and R, to produce more SdpI to offset SdpC, finally guaranteeing the subgroup alive, thereby delaying the spores production.We transfer SdpC which is fused by promoter SdpI/R into high copy plasmids in order to damage the balance of the system, thereby killing whole colony. When SdpC appears, SdpI on the membrane will bind free SdpC and adsorb SdpR to cease its inhibition against SdpI P/R, trying to produce more SdpI. At the same time, it will activate the promoter SdpR/I in our circuits and generate more SdpC. The system would fall into an infinite loop, and according to our modeling,the amount of SdpC increases beyond the ability of SdpI. Thus, the cells with protection mechanism will crack and die because of too much SdpC. All of them formed the killing device.</p><br />
<img src="https://static.igem.org/mediawiki/igem.org/5/5d/2013ustc-china_design11sdpABC.png" width="580" height="160"/><br />
<p>We Also designed a test circuit, which contains promotor grac and sdpABC only, aiming to determine the ability of SdpC.</p><br />
<br />
</div><br />
<br />
<br />
</div><br />
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</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/Project/DesignTeam:USTC CHINA/Project/Design2013-09-27T19:15:21Z<p>Surine: </p>
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<h1 style="font-size:40px;">Gene Circuit</h1><br />
<div class="basic-bar"><br />
<h2>Introduction</h2><br />
<p>Our T-vaccine consists four modules. Each of them carries a gene circuit independently. The first one expresses TD1-Antigen fusion protein, as the core of our vaccine. The second one expresses TD1-Anjuvant fusion proteins, which enhance the antigenicity. The third one is reporter, it notifies users whether the status of vaccine patch is all right and when they can stick the patches to arms. The last module is a kill switch,we designed a reliable suicide system in bacillus subtilis for the very first time in iGEM. So it would be OK if our users tear patches off and throw them away. We attribute different gene circuits to different bacteria(Modularization idea), by changing the ratio of each bacteria we could make it compatible with any vaccine.</p><br />
<img src="https://static.igem.org/mediawiki/2013/0/0a/2013ustc-chinaWorkflow.png" width="580" height="300" /><br />
</div><br />
<div class="basic-bar"><br />
<h2>Our Design</h2> <br />
<img src="https://static.igem.org/mediawiki/2013/4/4d/2013ustc-china_genecircuit.png" width="580" height="180"/><br />
<h3>1. Antigen</h3><br />
<p>As the main part of T-vaccine, TD-1 is fused with different kinds of antigens, so it can penetrate the skin and provoke immune responses. We have designed three kinds of vaccine, which are against hepatitis B, tuberculosis and anthrax. Our ELISA test had proved their antigenicity and mice test will figure it out if they bave immunogenicity,thereby proving that T-vaccine can be generally used. </p><br />
<br><br><br />
<h3>2. Adjuvant</h3><br />
<p>Similar with traditional injection vaccine, adjuvant should be added into vaccine to ensure immunity. We fused TD-1 with LTB. LT has adjuvant activity and can assist foreign antigen to induce the body to produce systemic immune response. The B submit of LTB protein has no toxic effect and it also works.</p><br />
<br><br><br />
<h3>3. TNFα</h3><br />
<p>TNFα can recruit Langerhans cell(LC) which work as antigen-presenting cells around epidermis, and improved LC could transmit into adjacent lymph node provoking the immune response. Because this circuit is quite similar with 1# and 2# ,while its effect might be difficult to certified, so we delayed the schedule of this part. 3# is the only circuit which has not been built or tested during the whole summer experiment.<br />
</div><br />
<br />
<br />
<br />
<div class="basic-bar"><br />
<h2>4. Reporter System</h2><br />
<p>To be more user-friendly, 4# contains a reporting system. After melting in water, the spores will germinate and express blue pigment protein (amilCP) to report the best using time. Meanwhile, 4# could also ensure biosafety, as 4# engineering bacteria can kill all the engineered bacteria after use.</p><br />
<h3>4.1 Reporter</h3><br />
<img src="https://static.igem.org/mediawiki/igem.org/a/aa/2013ustc-china_design111reporter.png" width="580" height="160"/><br />
<p>T-vaccine is also a user-friendly product. Our potential users are not medical professionals, they activate T-vaccine by exposing it to water, and the patch gives visualized signal to notify users whether the status of engineered bacteria is all right and when to stick the patch to arms. We achieve this simply with the regulation of promoter 43, which is recognized by sigma factor A. The activity of promoter 43 is maximal during the exponential growth phase. In other words, we expect engineering Bacillus subtilis to express blue pigment a few hours before expressing massive antigens and adjuvants. It is the perfect time.</p><br />
<br><br><br />
<h3>4.2 Kill Switch</h3><br />
<img src="https://static.igem.org/mediawiki/2013/f/f0/2013ustc-china_sdpABC.png" width="580" height="450"/><br />
<br><br><br />
<p>In B.subtilis, when it comes to the stationary phase, the environmental pressure increases and nutrition becomes limited, so B.subtilis begins to produce spore. Now the community will be divided into two different parts. One of them are trying to kill others to get enough nutrient, delaying the production of spores and achieving a competitive advantage. Killing is mediated by the exported toxic protein SdpC. SdpI will appear on the membrane surface to avoid itself from being damaged. SdpI could bind free SdpC and autopressor SdpR, to remove inhibition of SdpR against I and R, to produce more SdpI to offset SdpC, finally guaranteeing the subgroup alive, thereby delaying the spores production.We transfer SdpC which is fused by promoter SdpI/R into high copy plasmids in order to damage the balance of the system, thereby killing whole colony. When SdpC appears, SdpI on the membrane will bind free SdpC and adsorb SdpR to cease its inhibition against SdpI P/R, trying to produce more SdpI. At the same time, it will activate the promoter SdpR/I in our circuits and generate more SdpC. The system would fall into an infinite loop, and according to our modeling,the amount of SdpC increases beyond the ability of SdpI. Thus, the cells with protection mechanism will crack and die because of too much SdpC. All of them formed the killing device.</p><br />
<img src="https://static.igem.org/mediawiki/igem.org/5/5d/2013ustc-china_design11sdpABC.png" width="580" height="160"/><br />
<p>We Also designed a test circuit, which contains promotor grac and sdpABC only, aiming to determine the ability of SdpC.</p><br />
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<div></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/Project/Results/FurtherWorkTeam:USTC CHINA/Project/Results/FurtherWork2013-09-27T19:06:09Z<p>Surine: </p>
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<h1>Introduction<h1><br />
<p>According to our experiment result, we have proved the secretion and expression possibility of TD1-antigen, TD1-adjuvant, also the antigenicity of TD1-antigen after transdermal process. So in the following experiments, we decided to utilize Bacillus Subtillis WB800N as engineering bacteria, plus shuttle vector pHT43 as secretion vector to build the Bacillus Subtillis secretory expression system. On top of this, we have taken advantage of different kinds of TD1-antigen, testing HBsAg, PA, Ag85b that have been applied into market to check the universal property of TD1-antigen. Besides, reporters that are essential during reality application have been found to make the final circuit come true.<p><br />
<br />
<br />
<img src="https://static.igem.org/mediawiki/2013/c/cb/Further_work.png"width="580" height="250" /><br />
<div><h2>ONE:Expression of TD1-"X" in WB800N</h2></div><br />
<div><h3>1.Expression of TD1-GFP</h3></div> <br />
<img src="https://static.igem.org/mediawiki/2013/c/c3/2013ustc-chinaWB800N-GFP.png" width="580" height="420" /><br />
<br />
<div><h3>2.Expression of TD1-antigen/adjuvant</h3></div><br />
<div><p>In order to realize the secretory expression in Bacillus Subtillis, we inserted signal peptide between promoter and the TD1-Antigen/adjuvant sequence to secrete our recombinant protein. GFP has been chosen to check whether the reliability of this circuit. After large amounts of experiments, GFP has finally been found via fluorescence microscope. </p></div><br />
<img src="https://static.igem.org/mediawiki/2013/f/f0/Pctc-sp-td1-antigenadjuvent.png"width="580" height="120" /><br />
<br />
<div><p>Then, three kinds of recombinant antigen, TD1-HBsAg, TD1-PA, TD1-Ag85b, and recombinant immunologic adjuvant were designed. The earliest successful one TD1-LTB protein, can not only be observed clear stripes in SDS-PAGE, but also be proved according to HPLC-MS.</p></div><br />
<img src="https://static.igem.org/mediawiki/igem.org/f/fd/20-%E6%9E%AF%E8%8D%89%E8%9B%8B%E7%99%BD%E8%A1%A8%E8%BE%BE.jpg" width="580" height="300"><br />
<div class="atfigure" align="center" style="width:580px;font-size:14px;">Fig4. SDS PAGE shows the expression of LTB in WB800N</div><br />
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<h1>Introduction<h1><br />
<p>According to our experiment result, we have proved the secretion and expression possibility of TD1-antigen, TD1-adjuvant, also the antigenicity of TD1-antigen after transdermal process. So in the following experiments, we decided to utilize Bacillus Subtillis WB800N as engineering bacteria, plus shuttle vector pHT43 as secretion vector to build the Bacillus Subtillis secretory expression system. On top of this, we have taken advantage of different kinds of TD1-antigen, testing HBsAg, PA, Ag85b that have been applied into market to check the universal property of TD1-antigen. Besides, reporters that are essential during reality application have been found to make the final circuit come true.<p><br />
<br />
<br />
<img src="https://static.igem.org/mediawiki/2013/c/cb/Further_work.png"width="580" height="250" /><br />
<div><h2>ONE:Expression of TD1-"X" in WB800N</h2></div><br />
<div><h3>1.Expression of TD1-GFP</h3></div> <br />
<img src="https://static.igem.org/mediawiki/2013/c/c3/2013ustc-chinaWB800N-GFP.png" width="580" height="120" /><br />
<br />
<div><h3>2.Expression of TD1-antigen/adjuvant</h3></div><br />
<div><p>In order to realize the secretory expression in Bacillus Subtillis, we inserted signal peptide between promoter and the TD1-Antigen/adjuvant sequence to secrete our recombinant protein. GFP has been chosen to check whether the reliability of this circuit. After large amounts of experiments, GFP has finally been found via fluorescence microscope. </p></div><br />
<img src="https://static.igem.org/mediawiki/2013/f/f0/Pctc-sp-td1-antigenadjuvent.png"width="580" height="120" /><br />
<br />
<div><p>Then, three kinds of recombinant antigen, TD1-HBsAg, TD1-PA, TD1-Ag85b, and recombinant immunologic adjuvant were designed. The earliest successful one TD1-LTB protein, can not only be observed clear stripes in SDS-PAGE, but also be proved according to HPLC-MS.</p></div><br />
<img src="https://static.igem.org/mediawiki/igem.org/f/fd/20-%E6%9E%AF%E8%8D%89%E8%9B%8B%E7%99%BD%E8%A1%A8%E8%BE%BE.jpg" width="580" height="300"><br />
<div class="atfigure" align="center" style="width:580px;font-size:14px;">Fig4. SDS PAGE shows the expression of LTB in WB800N</div><br />
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</html></div>Surinehttp://2013.igem.org/File:2013ustc-chinaWB800N-GFP.pngFile:2013ustc-chinaWB800N-GFP.png2013-09-27T19:03:41Z<p>Surine: uploaded a new version of &quot;File:2013ustc-chinaWB800N-GFP.png&quot;</p>
<hr />
<div></div>Surinehttp://2013.igem.org/File:2013ustc-chinaWB800N-GFP.pngFile:2013ustc-chinaWB800N-GFP.png2013-09-27T19:01:41Z<p>Surine: uploaded a new version of &quot;File:2013ustc-chinaWB800N-GFP.png&quot;</p>
<hr />
<div></div>Surinehttp://2013.igem.org/File:2013ustc-chinaWB800N-GFP.pngFile:2013ustc-chinaWB800N-GFP.png2013-09-27T18:59:32Z<p>Surine: </p>
<hr />
<div></div>Surinehttp://2013.igem.org/File:2013ustc-china_Needles.pngFile:2013ustc-china Needles.png2013-09-27T18:29:26Z<p>Surine: uploaded a new version of &quot;File:2013ustc-china Needles.png&quot;</p>
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<div></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/Project/OverviewTeam:USTC CHINA/Project/Overview2013-09-27T18:26:11Z<p>Surine: </p>
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<h1>Overview</h1><br />
<p>In our world, billions of people are suffering from contagions. However, only a fraction of contagions can be prevented by the existing vaccines. The disadvantages of traditional vaccines, which are bothered to produced and purified and has strict requirements about temperature, limit their usage especially in developing countries. </p><br />
<div align="center"><img src="https://static.igem.org/mediawiki/igem.org/e/ec/2013ustc-china_traditional_vaccine_transport.jpg" width="400" height="300" /><br />
<div class="atfigure" align="center" style="width:400px;font-size:14px;">Fig1. Difficult shipping in remote areas </div></div><br />
<p>This year, our project focused on a revolutionary vaccine delivery. We bring a fresh Medication into the world which is an in situ expression system. Our practice of this concept is a biological transdermal vaccine patch called T-vaccine. We chose Bacillus subtilis as chassis to establish the band-aid secreting fresh vaccines. The new vaccine consist of four engineering B.subtilis, each of which carried a gene circuit independently.</p><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2013/archive/e/ed/20130923171924!2013igemustc_Standardization.png" width="400" height="350" /><br />
<div class="atfigure" align="center" style="width:400px;font-size:14px;">Fig2. block-based design </div></div><br />
<p>With an excellent transdermal peptide TD1, three of the engineering B.subtilis could express a series of fusion proteins (the antigen and two kinds of adjuvants) which could penetrate the skin and work as traditional vaccine molecules. The fourth bacteria are our “reporter” which would notify users when the band-aid is working well and the patch can be pasted . Moreover, we designed a reliable suicide system in Bacillus subtilis to ensure the biosafety.</p><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2013/9/91/2013ustc-china_Needles.png" width="400" height="350" /><br />
<div class="atfigure" align="center" style="width:400px;font-size:14px;">Fig3. No Needle </div></div><br />
<p>T-vaccine has great advantages in transportation which can be stored from minus 20 to 60 Celsius. With T-vaccine, we can reach every remote corner of the world and help eliminate contagions from our world. Also it is proven that transdermal vaccine is an efficient and effective way for a variety of pathogens such as: tuberculosis, anthrax, hepatitis B and so on. Last but not least, we have created a world free from needles. Given all these advantages, T-vaccine is expected to be a promising vaccine research and development direction.</p> <br />
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<h1>Overview</h1><br />
<p>In our world, billions of people are suffering from contagions. However, only a fraction of contagions can be prevented by the existing vaccines. The disadvantages of traditional vaccines, which are bothered to produced and purified and has strict requirements about temperature, limit their usage especially in developing countries. </p><br />
<div align="center"><img src="https://static.igem.org/mediawiki/igem.org/e/ec/2013ustc-china_traditional_vaccine_transport.jpg" width="400" height="300" /><br />
<div class="atfigure" align="center" style="width:400px;font-size:14px;">Fig1. Difficult shipping in remote areas </div></div><br />
<p>This year, our project focused on a revolutionary vaccine delivery. We bring a fresh Medication into the world which is an in situ expression system. Our practice of this concept is a biological transdermal vaccine patch called T-vaccine. We chose Bacillus subtilis as chassis to establish the band-aid secreting fresh vaccines. The new vaccine consist of four engineering B.subtilis, each of which carried a gene circuit independently.</p><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2013/archive/e/ed/20130923171924!2013igemustc_Standardization.png" width="400" height="350" /><br />
<div class="atfigure" align="center" style="width:400px;font-size:14px;">Fig2. block-based design </div></div><br />
<p>With an excellent transdermal peptide TD1, three of the engineering B.subtilis could express a series of fusion proteins (the antigen and two kinds of adjuvants) which could penetrate the skin and work as traditional vaccine molecules. The fourth bacteria are our “reporter” which would notify users when the band-aid is working well and the patch can be pasted . Moreover, we designed a reliable suicide system in Bacillus subtilis to ensure the biosafety.</p><br />
<div align="center"><img src=https://static.igem.org/mediawiki/2013/9/91/2013ustc-china_Needles.png"" width="400" height="350" /><br />
<div class="atfigure" align="center" style="width:400px;font-size:14px;">Fig3. No Needle </div></div><br />
<p>T-vaccine has great advantages in transportation which can be stored from minus 20 to 60 Celsius. With T-vaccine, we can reach every remote corner of the world and help eliminate contagions from our world. Also it is proven that transdermal vaccine is an efficient and effective way for a variety of pathogens such as: tuberculosis, anthrax, hepatitis B and so on. Last but not least, we have created a world free from needles. Given all these advantages, T-vaccine is expected to be a promising vaccine research and development direction.</p> <br />
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<h1>Attributions</h1><br />
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<p> <br />
In the last two months,thanks to our devotion and the support of various professors and scientific institutions ,our team achieved proud results.<br><br />
In the summer vacation,we used the gene material donated by scientific institutions and individuals to standardize and splice.we standardized and spliced transdermal peptide and gene circuits consists of various fusion protein of antigen.Meanwhile,we expressed TD1-GFP、TD1-HBsAg、TD1-LTB and all of expression of them was verified by in the E.coli. TD1-PA、TD1-HBsAg、TD1-Ag85b and TD1-LTB constructed circuits of the secretion vector,WB800N.Firstly,the transdermal experiment outside human body succeeded.Secondly,we also try to do experiment in living mice.Generally,we construct 16 new parts,we set a foundation for toolkit.Also our standardized BBa_K1074006 set a frame for other secretion expression researchers,theoretically users can use TD1 to merge any protein,to made the target protein have the function of secretion and transdermal permeation.<br><br />
<br> <br />
Thanks to the following individuals and institutions:<br><br />
1.Gene donation<br><br />
Standardized antigen:Chinese Centre for Disease<br><br />
TD-1 short chain poly peptide:Professor Longping Wen<br><br />
2.Theoretically support:<br><br />
Consultation of Bacillus subtilis:Professor Xin Run,Nanjing Agriculture University<br><br />
Consultation of transdermal experiment:Professor Longping Wen<br><br />
Training of mice experiment:Professor Ting Yue<br><br />
<br />
3.Preliminary Partner:Hualan Biology Co.,ltd.<br><br />
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<h1>Attributions</h1><br />
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<p> <br />
In the last two months,thanks to our devotion and the support of various professors and scientific institutions ,our team achieved proud results.<br><br />
In the summer vacation,we used the gene material donated by scientific institutions and individuals to standardize and splice.we standardized and spliced transdermal peptide and gene circuits consists of various fusion protein of antigen.Meanwhile,we expressed TD1-GFP、TD1-HBsAg、TD1-LTB and all of expression of them was verified by in the E.coli. TD1-PA、TD1-HBsAg、TD1-Ag85b and TD1-LTB constructed circuits of the secretion vector,WB800N.Firstly,the transdermal experiment outside human body succeeded.Secondly,we also try to do experiment in living mice.Generally,we construct 16 new parts,we set a foundation for toolkit.Also our standardized BBa_K1074006 set a frame for other secretion expression researchers,theoretically users can use TD1 to merge any protein,to made the target protein have the function of secretion and transdermal permeation.<br><br />
<br> <br />
Thanks to the following individuals and institutions:<br><br />
1.Gene donation<br><br />
Standardized antigen:Chinese Centre for Disease<br><br />
TD-1 short chain poly peptide:Professor Longping Wen<br><br />
2.Theoretically support:<br><br />
Consultation of Bacillus subtilis:Professor Xin Run,Nanjing Agriculture University<br><br />
Consultation of transdermal experiment:Professor Longping Wen<br><br />
Training of mice experiment:Professor Ting Yue<br><br />
<br />
3.Preliminary Partner:Hualan Biology Co.,ltd.<br><br />
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<h2>Project Safety</h2><br />
<h3>Would any of your project ideas raise safety issues in terms of researcher safety,public safety or environmental safety?</h3><br />
<p>To ensure biosafety, we design a <a href="https://2013.igem.org/Team:USTC_CHINA/Project/Design" target="_blank" >kill switch</a> in Bacillus for the first time in iGEM. The very type B.subtilis, which contains kill switch circuit, can elimate their siblings and commit suicide when the switch is activated, and thus prevent gene contamination and raise project safty in terms of researcher safety, public safety and environmental safety.</p><br />
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<h2>Parts Safety</h2><br />
<h3>Do any of the new BioBrick part (or devices) that you made this year raise any safety issues?</h3><br />
<p>The only safety issue associated with our new BioBrick parts is researcher safety in testing the parts. We've revised a strict Lab operation standard to raise safty.<br><br />
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<h2>Safty Supervision</h2><br />
<h3>Is there a local biosafety group, committee, or review board at your institution? </h3><br />
<p>Yes.Supervised by biosafety committee,we accepted their advice and abandoned intention to use HBV genome as template to amplify HBsAg antigen. With their help,we obtained plasmid fused HBsAg antigen eventually.</p><br />
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<h1>Introduction</h1><br />
To be more user-friendly, 4# circuit contains a reporting system. After melting in water, the spores germinate and express blue pigment protein to report the best using time.</br><br />
Meanwhile, 4# circuit could also ensure biosafety. Because other circuit do not have self-killing device, 4# engineering bacterial should kill all the bacterial after using.</br><br />
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<h1>Designing of the suicide system</h1><br />
We design a circuit of killing switch based on its endogenous genetic system.</br><br />
In B.subtilis, when it comes to the stationary phase, the environmental pressure increases and nutrition becomes limited, so B begin to produce spores. Now the community will be divided into two different parts. One of them are trying to kill others to get enough nutrient , delaying the production of spores and achieving a competitive advantage. Killing is mediated by the exported toxic protein SdpC. SdpI will appear on the membrane surface to avoid itself from being damaged. SdpI could bind free SdpC and autopressor SdpR, to remove SdpR’s inhibition against I and R, to produce more SdpI to offset SdpC, finally guaranteeing the subgroup alive, thereby delaying the spores production.</br><br />
<img src="https://static.igem.org/mediawiki/2013/2/2b/Reporter_3.png" width="500" height="350"><br />
<p>We transfer SdpC which is fused by promoter SdpI/R into high copy plasmids in order to damage the balance of the system, thereby killing whole colony. When SdpC appears, SdpI on the membrane will bind free SdpC and adsorb SdpR to cease its inhibition against SdpI P/R, trying to produce more SdpI. At the same time, it will activate the promoter SdpR/I in our circuits and generate more SdpC.The system would fall into an infinite loop, and according to our modeling ,the amount of SdpC increases beyond the ability of SdpI.Thus,the cells with protection mechanism will crack and die because of too much SdpC. All above forms the killing device.<br />
We Also designed a test circuit,which contains promotor grac and sdpABC only,aiming to determine the ability of SdpC.</p><br />
<br />
<div></br></br><br />
There are both positive and negative feedback loops in this process. On the one hand, SdpI is unable to sequestrate the autorepressor, SdpR, until it captures the toxin, SdpC. The accumulation of SdpC will thus facilitate SdpI to capture more SdpR and thereby relieve the repression of SdpR, stimulating the expression of itself. This is the positive feedback loop which leads to the increasing accumulation of SdpC and finally the death of the bacteria. On the other hand, the removal of SdpR also enhance the expression of SdpI and accelerate the sequestration of SdpC, which forms a negative feedback loop whose effects contradict the positive feedback loop. However, since the copy number of SdpC is much higher, it is believed that the positive loop is strong enough to outweigh the negative one, which guarantees this mechanism will finally leads to collapse instead of equilibrium.<br />
</div><br />
<div><br />
<h1>The ODE model of singular cells</h1><br />
There is no denying fact that the essential goal of engineered bacterias who carry this so called “suicide” locus itself is to kill their siblings rather than themselves to ensure the survival of themselves. Surly they can kill their siblings, but can they finally eliminate themselves, as we expects? The trivial experiment protocol and huge uncertainty had put off our experiment, and as expected, we failed to achieve the construction of complete reporter system in our laboratory. Fortunately, we could resort to mathematical models to verify the validity of this locus theoretically. <br />
There are six independent variables in individual cells, and the theoretically if the initial conditions are fixed, all of them will be the univariate functions of time. The following table illustrates the mark and meaning of each variable:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub></td><br />
<td >Mole number of free SdpI in cytoplasm</a>.</td><br />
</tr><br />
<tr><br />
<td >I<sub>m</sub> </td><br />
<td >Mole number of SdpI in the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >C<sub>f</sub></td><br />
<td >Mole number of free SdpC in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >C<sub>i</sub></td><br />
<td >Mole number of SdpC captured by SdpI.</td><br />
</tr><br />
<tr><br />
<td >R<sub>f</sub></td><br />
<td >Mole number of free SdpR in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >R<sub>i</sub> </td><br />
<td >Mole number of SdpR captured by SdpI</td><br />
</tr><br />
</table><br />
</br></br><br />
<div>To construct reasonable ordinary differential equation (ODE) model to describe and predict the operation of the suicide system, we followed the law of mass action, one basic law of chemistry and biology.</br><br />
Taken as a statement about kinetics, the law states that the rate of an elementary reaction (a reaction that proceeds through only one transition state, which is one mechanistic step) is proportional to the product of the concentrations of the participating molecules. In modern chemistry this is derived using statistical mechanics. Despite the complicated chemical reactions involved in the process of transcription and translation, it is common and logically sound to view the expression of one particular gene as an elementary reaction and assume the repression effects of the protein itself encodes and the repressor are both linear.</br><br />
According to the law of mass action, we got six independent differential equation of the variables:</br><br />
<img src="https://static.igem.org/mediawiki/2013/e/e8/For1%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/2/23/For2%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/4/48/For3%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/7/71/For4%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/3/31/For5%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/c/cb/For6%281%29.png"></br></br></br><br />
The following table explain the constants in the above ODE groups:</br></br></div><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub> </td><br />
<td >The maximal number of SdpI than can be fixed on the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >k<sub>0</sub></td><br />
<td >Constant describes the normal expression rate of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>1</sub> </td><br />
<td >Constant describes the self-repression effects of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>2</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpC.</a></td><br />
</tr><br />
<tr><br />
<td >k<sub>3</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>4</sub> </td><br />
<td >Constant describes the normal expression rate of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>5</sub> </td><br />
<td >Constant describes the self-repression effects of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>6</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>7</sub> </td><br />
<td >Constant describes the normal expression rate of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>8</sub> </td><br />
<td >Constant describes the self-repression effects of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>9</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>10</sub> </td><br />
<td >Constant describes the rate of SdpI binding to the cell membrane</td><br />
</tr><br />
</table><br />
<br />
<br />
<br />
<br />
</div><br />
<div class="clear"></div><br />
<div><br />
<h2>Discussions on the constants</h2><br />
All the constants given above is steady and theoretically measurable when all the conditions are constant. For example, we could measure k<sub>0</sub> by constructing a new engineered bacteria, which contains the gene encoding SdpC and marker gene alone and observing the influence of the concentration of SdpC on its expression. Yet any modification on genome is notoriously time-consuming, which inhibited us from measuring them in person. We also looked up oceans of papers to confer their approximate ranges, but almost all papers are too fragmental to afford any valid information. Therefore, we decided to assume all these constant according to our limited information and make a qualitative analysis instead of quantifiable analysis. All units and dimensions were temporarily ignored. In other words, our model aims at justifying the validity of this suicide mechanism rather than predicting the exact time or any other parameters of the system.<br />
<br />
Despite the fact that we have hardly any accurate data on these constants, there are some limitations that we extrapolated from known information before we further explore this model:</br><br />
<ol><br />
<li>k<sub>0</sub>>>k<sub>4</sub>≈k<sub>7</sub>: k<sub>0</sub>,k<sub>4</sub> and k<sub>7</sub> represent the normal expression rate of SdpC, SdpR and SdpC separately, and the copy number of SdpC is much larger than that of SdpR and SdpI, whereas the value of the latter two is approximately equal;</li><br />
<li>k<sub>2</sub>>>k<sub>9</sub>: the existence of free SdpR represses the expression of both SdpI and SdpC, and similarly, since the copy number of SdpC is much higher, we expected the repression effect was stronger accordingly;</li><br />
<li>k<sub>10</sub>>>k<sub>3</sub>,k<sub>6</sub>:it is hard to predict the value of k<sub>3</sub> and k<sub>8</sub>, yet we suppose both of them is much smaller than k<sub>10</sub> because SdpI is a kind of membrane protein inherently, and rarely exists as free protein</li><br />
<li>The primary values of all the six variables are very small or strictly zero. We expect it as the most logical initial status. If the primary value of any variable is relatively large, the suicide mechanism may not run normally</li><br />
</ol><br />
</br></br><br />
<h2>Stimulation and discussion</h2><br />
<br />
Simple and rough as the above model is, it does theoretically sound. To test the validity of this model, we first tried to get analytic solution of the ODE set. If this analytic solution exists, we could further investigate the interaction among those variables, and draw some phase planes to get accurate and mathematically perfect description of this model. Unfortunately but expectedly, the existence of analytic solution was negated by MATLAB, and we had to assume groups of values for these constants in advance and analyze the arithmetic solutions instead. These arithmetic solutions not only justified this mechanism is effective enough to commit cell suicide but also indicated some unexpected, or even weird results that beyond our wildest imagination. There are two possibility account for the unexpected results: our model is too rough to include some assignable factor; or there are some implicit but objective limitation inside model, which may be substantiate by later experiments or papers.</br><br />
When we explored the arithmetic solutions of this ODE set, we received nearly one hundred warnings from MATLAB and for many times our most powerful computer ran out of its 8GB memory, but sometimes we can receive the solution within seconds. We had adjusted our parameters for several times before we got our first solution. Here is the values of parameters for this group, and the graph of arithmetic solutions is also given:</br><br />
<div style="margin-top:20px;"><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>50</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table><br />
</div></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/2/26/Suicide1.png"></br></br><br />
At the first glance this graph seemed fine. Initially the concentration of SdpC decreased slightly due to the capturing of SdpI and the repression of float SdpR, but gradually the positive feedback loop works, and C<sub>f</sub> increases rapidly. But when we turned our attention to the curves of other parameters, things seemed not so perfect:</br><br />
<div><img class="linegraph" src="https://static.igem.org/mediawiki/2013/c/cf/Suicide2.png"></br></br><br />
The curve of I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub> contradicted our common sense severely. First, I<sub>m</sub>>C<sub>i</sub>>R<sub>i</sub> is expected to be tenable all the time, which precludes the intersects among the three curves; Second, there is no mechanism in this system that could decrease their concentration, and all of them are increasing function; Third and most serious, never will them be negative, as they represent the concentration of real substances.<br />
<br />
Then we adjusted the parameters slightly for several times. To eliminate those absurd curves, we reconsidered some assumptions.<br />
Here we listed another representative group of parament values and relative graph:</div></br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/7/7d/Suicide3.png/800px-Suicide3.png"></br></br><br />
In this group, we gave up one former assumption and set k<sub>2</sub> equal to k<sub>9</sub>. We also gave positive values to I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub>, which were considered to be zero at first. And by groups of stimulations we realized the value of k<sub>2</sub> does matter, as the derivative of C<sub>f</sub> only increased slightly as k<sub>2</sub> lowers, and the positive values failed to avoid the weird phenomenon in the latter three curves.<br />
We also found that however we adjusted the primary value of I<sub>f</sub> and other parameters, If dropped into approximately zero extremely rapidly at the initial stage and remained balanced, which might account for why the derivatives of the latter curves were abnormally negative. Thus we modified another assumption and increased k<sub>7</sub>. Here is another group of values and corresponding graph:</br></br><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/1/15/Suicide4.png/800px-Suicide4.png" style="width:600; height:400;"></br></br><br />
Although the derivative of Im is not seriously positive constantly, the three latter curves seemed much more reasonable. Hence, we extrapolated although SdpI and SdpR share the same promoter, the expression of SdpI must much faster than SdpR to ensure successful “suicide.” Additionally, the increase of k<sub>7</sub> also represses SdpC, and hence the copy number of SdpC must be larger.<br />
We kept all other parameters constant and gradually augmented k<sub>0</sub>. The larger k<sub>0</sub>, the more perfect the curve seemed, and here are the values table and graph where k<sub>0</sub> equals 400, 80 times larger than k<sub>4</sub>.</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>400</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/3/33/Suicide5.png/800px-Suicide5.png"></br></br><br />
Take the curve of C<sub>f</sub> and R<sub>f</sub> separately, the curves seemed more perfect:</br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/9/9f/Suicide6.png"></br></br><br />
<br />
In wild bacteria who are unable to produced SdpC, naturally k<sub>0</sub> equals zero. We expected C<sub>f</sub> would decreased gradually and finally approximate zero, and here are the corresponding table and graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>30</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/1/19/Suicide7.png"></br></br><br />
Wired but not surprising, there were intersects among the latter three curve, and C<sub>f</sub> decreases continually when it is negative. We continued to try groups of these parameters, and this is the best one where we increased the primary concentration of SdpC and the normal expression rate of SdpI.<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/8/8e/Suicide8.png"></br></br><br />
The curve of C<sub>f</sub> and R<sub>f</sub> alone:<br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/e/eb/Suicide9.png"></br></br><br />
In spite of minimal abnormal phenomenon (C<sub>f</sub> was negative in later stage), this graph roughly testified that in wild bacterial the concentration of float SdpC will drop to nearly zero quickly.<br />
In sum, the ODE model of singular cells indicate following results:</br><br />
<ol><br />
<li>The character of free SdpC is most affected by k<sub>0</sub>, if the copy number of SdpC is large enough, it is theoretically reasonable to commit suicide;</li><br />
<li>The influence of the value of Imax and k<sub>2</sub> is much limited;</li><br />
<li>The amount of free SdpI is always near zero;</li><br />
<li>SdpC will not increase limitlessly however we transform parameters;</li><br />
<li>To ensure the success of suicide, it is required k<sub>0</sub>>>k<sub>4</sub>>>k<sub>7</sub>;</li><br />
The last conclusion was our biggest windfall, and we have verified the validity of this suicide mechanism in math. On the one hand, if further experiments proven #4 engineered bacteria will kill both siblings and themselves, it is highly like that the expression rate SdpI is much larger than SdpR even if they share the same promoter; on the other hand, if #4 engineered bacteria are not able to commit suicide, we can try to boost the expression of SdpI to adjust the bacteria.<br />
</ol><br />
<br />
<div><br />
<h1>Discussion on colonies</h1><br />
<br />
In reality, the engineered bacteria aims at killing its siblings instead of itself, and at first almost all toxin SdpC will be secreted outside the bacteria. Assume the diffusion of toxin among cells comply with diffusion law, that is, the diffusion rate is proportionate with the gradient of concentration. Further assume the death concentration of SdpC is same to all bacteria expect those who contain this locus, the average life expectancy is bacteria will hinge on the rate and distribution of engineered bacteria, and the distribution of life expectancy of bacteria is similar to that of average free path of gas molecules.<br />
As long the coefficient of diffusion is large enough, any engineered bacterias, no matter how few, is adequate to devastate the whole colony. Alike to the average free path of thin gas, the average suicide time of the whole reporter system is inversely proportional with the square root of the rate of engineer bacteria containing this locus.<br />
</div><br />
<br />
<h1>References</h1><br />
Parallel pathways of repression and antirepression governing the transition to stationary phase in Bacillus subtilis<br />
AV Banse, A Chastanet, L Rahn-Lee…,PNAS ,2008 </br><br />
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<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a>&gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></div></div><br />
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<h1>Introduction</h1><br />
To be more user-friendly, 4# circuit contains a reporting system. After melting in water, the spores germinate and express blue pigment protein to report the best using time.</br><br />
Meanwhile, 4# circuit could also ensure biosafety. Because other circuit do not have self-killing device, 4# engineering bacterial should kill all the bacterial after using.</br><br />
<br />
</div><br />
<br />
<div><br />
<h1>Designing of the suicide system</h1><br />
We design a circuit of killing switch based on its endogenous genetic system.</br><br />
In B.subtilis, when it comes to the stationary phase, the environmental pressure increases and nutrition becomes limited, so B begin to produce spores. Now the community will be divided into two different parts. One of them are trying to kill others to get enough nutrient , delaying the production of spores and achieving a competitive advantage. Killing is mediated by the exported toxic protein SdpC. SdpI will appear on the membrane surface to avoid itself from being damaged. SdpI could bind free SdpC and autopressor SdpR, to remove SdpR’s inhibition against I and R, to produce more SdpI to offset SdpC, finally guaranteeing the subgroup alive, thereby delaying the spores production.</br><br />
<img src="https://static.igem.org/mediawiki/2013/2/2b/Reporter_3.png" width="500" height="350"><br />
<p>We transfer SdpC which is fused by promoter SdpI/R into high copy plasmids in order to damage the balance of the system, thereby killing whole colony. When SdpC appears, SdpI on the membrane will bind free SdpC and adsorb SdpR to cease its inhibition against SdpI P/R, trying to produce more SdpI. At the same time, it will activate the promoter SdpR/I in our circuits and generate more SdpC.The system would fall into an infinite loop, and according to our modeling ,the amount of SdpC increases beyond the ability of SdpI.Thus,the cells with protection mechanism will crack and die because of too much SdpC. All above forms the killing device.<br />
We Also designed a test circuit,which contains promotor grac and sdpABC only,aiming to determine the ability of SdpC.</p><br />
<br />
<div></br></br><br />
There are both positive and negative feedback loops in this process. On the one hand, SdpI is unable to sequestrate the autorepressor, SdpR, until it captures the toxin, SdpC. The accumulation of SdpC will thus facilitate SdpI to capture more SdpR and thereby relieve the repression of SdpR, stimulating the expression of itself. This is the positive feedback loop which leads to the increasing accumulation of SdpC and finally the death of the bacteria. On the other hand, the removal of SdpR also enhance the expression of SdpI and accelerate the sequestration of SdpC, which forms a negative feedback loop whose effects contradict the positive feedback loop. However, since the copy number of SdpC is much higher, it is believed that the positive loop is strong enough to outweigh the negative one, which guarantees this mechanism will finally leads to collapse instead of equilibrium.<br />
</div><br />
<div><br />
<h1>The ODE model of singular cells</h1><br />
There is no denying fact that the essential goal of engineered bacterias who carry this so called “suicide” locus itself is to kill their siblings rather than themselves to ensure the survival of themselves. Surly they can kill their siblings, but can they finally eliminate themselves, as we expects? The trivial experiment protocol and huge uncertainty had put off our experiment, and as expected, we failed to achieve the construction of complete reporter system in our laboratory. Fortunately, we could resort to mathematical models to verify the validity of this locus theoretically. <br />
There are six independent variables in individual cells, and the theoretically if the initial conditions are fixed, all of them will be the univariate functions of time. The following table illustrates the mark and meaning of each variable:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub></td><br />
<td >Mole number of free SdpI in cytoplasm</a>.</td><br />
</tr><br />
<tr><br />
<td >I<sub>m</sub> </td><br />
<td >Mole number of SdpI in the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >C<sub>f</sub></td><br />
<td >Mole number of free SdpC in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >C<sub>i</sub></td><br />
<td >Mole number of SdpC captured by SdpI.</td><br />
</tr><br />
<tr><br />
<td >R<sub>f</sub></td><br />
<td >Mole number of free SdpR in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >R<sub>i</sub> </td><br />
<td >Mole number of SdpR captured by SdpI</td><br />
</tr><br />
</table><br />
</br></br><br />
<div>To construct reasonable ordinary differential equation (ODE) model to describe and predict the operation of the suicide system, we followed the law of mass action, one basic law of chemistry and biology.</br><br />
Taken as a statement about kinetics, the law states that the rate of an elementary reaction (a reaction that proceeds through only one transition state, which is one mechanistic step) is proportional to the product of the concentrations of the participating molecules. In modern chemistry this is derived using statistical mechanics. Despite the complicated chemical reactions involved in the process of transcription and translation, it is common and logically sound to view the expression of one particular gene as an elementary reaction and assume the repression effects of the protein itself encodes and the repressor are both linear.</br><br />
According to the law of mass action, we got six independent differential equation of the variables:</br><br />
<img src="https://static.igem.org/mediawiki/2013/e/e8/For1%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/2/23/For2%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/4/48/For3%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/7/71/For4%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/3/31/For5%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/c/cb/For6%281%29.png"></br></br></br><br />
The following table explain the constants in the above ODE groups:</br></br></div><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub> </td><br />
<td >The maximal number of SdpI than can be fixed on the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >k<sub>0</sub></td><br />
<td >Constant describes the normal expression rate of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>1</sub> </td><br />
<td >Constant describes the self-repression effects of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>2</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpC.</a></td><br />
</tr><br />
<tr><br />
<td >k<sub>3</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>4</sub> </td><br />
<td >Constant describes the normal expression rate of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>5</sub> </td><br />
<td >Constant describes the self-repression effects of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>6</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>7</sub> </td><br />
<td >Constant describes the normal expression rate of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>8</sub> </td><br />
<td >Constant describes the self-repression effects of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>9</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>10</sub> </td><br />
<td >Constant describes the rate of SdpI binding to the cell membrane</td><br />
</tr><br />
</table><br />
<br />
<br />
<br />
<br />
</div><br />
<div class="clear"></div><br />
<div><br />
<h2>Discussions on the constants</h2><br />
All the constants given above is steady and theoretically measurable when all the conditions are constant. For example, we could measure k<sub>0</sub> by constructing a new engineered bacteria, which contains the gene encoding SdpC and marker gene alone and observing the influence of the concentration of SdpC on its expression. Yet any modification on genome is notoriously time-consuming, which inhibited us from measuring them in person. We also looked up oceans of papers to confer their approximate ranges, but almost all papers are too fragmental to afford any valid information. Therefore, we decided to assume all these constant according to our limited information and make a qualitative analysis instead of quantifiable analysis. All units and dimensions were temporarily ignored. In other words, our model aims at justifying the validity of this suicide mechanism rather than predicting the exact time or any other parameters of the system.<br />
<br />
Despite the fact that we have hardly any accurate data on these constants, there are some limitations that we extrapolated from known information before we further explore this model:</br><br />
<ol><br />
<li>k<sub>0</sub>>>k<sub>4</sub>≈k<sub>7</sub>: k<sub>0</sub>,k<sub>4</sub> and k<sub>7</sub> represent the normal expression rate of SdpC, SdpR and SdpC separately, and the copy number of SdpC is much larger than that of SdpR and SdpI, whereas the value of the latter two is approximately equal;</li><br />
<li>k<sub>2</sub>>>k<sub>9</sub>: the existence of free SdpR represses the expression of both SdpI and SdpC, and similarly, since the copy number of SdpC is much higher, we expected the repression effect was stronger accordingly;</li><br />
<li>k<sub>10</sub>>>k<sub>3</sub>,k<sub>6</sub>:it is hard to predict the value of k<sub>3</sub> and k<sub>8</sub>, yet we suppose both of them is much smaller than k<sub>10</sub> because SdpI is a kind of membrane protein inherently, and rarely exists as free protein</li><br />
<li>The primary values of all the six variables are very small or strictly zero. We expect it as the most logical initial status. If the primary value of any variable is relatively large, the suicide mechanism may not run normally</li><br />
</ol><br />
</br></br><br />
<h2>Stimulation and discussion</h2><br />
<br />
Simple and rough as the above model is, it does theoretically sound. To test the validity of this model, we first tried to get analytic solution of the ODE set. If this analytic solution exists, we could further investigate the interaction among those variables, and draw some phase planes to get accurate and mathematically perfect description of this model. Unfortunately but expectedly, the existence of analytic solution was negated by MATLAB, and we had to assume groups of values for these constants in advance and analyze the arithmetic solutions instead. These arithmetic solutions not only justified this mechanism is effective enough to commit cell suicide but also indicated some unexpected, or even weird results that beyond our wildest imagination. There are two possibility account for the unexpected results: our model is too rough to include some assignable factor; or there are some implicit but objective limitation inside model, which may be substantiate by later experiments or papers.</br><br />
When we explored the arithmetic solutions of this ODE set, we received nearly one hundred warnings from MATLAB and for many times our most powerful computer ran out of its 8GB memory, but sometimes we can receive the solution within seconds. We had adjusted our parameters for several times before we got our first solution. Here is the values of parameters for this group, and the graph of arithmetic solutions is also given:</br><br />
<div style="margin-top:20px;"><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>50</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table><br />
</div></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/2/26/Suicide1.png"></br></br><br />
At the first glance this graph seemed fine. Initially the concentration of SdpC decreased slightly due to the capturing of SdpI and the repression of float SdpR, but gradually the positive feedback loop works, and C<sub>f</sub> increases rapidly. But when we turned our attention to the curves of other parameters, things seemed not so perfect:</br><br />
<div><img class="linegraph" src="https://static.igem.org/mediawiki/2013/c/cf/Suicide2.png"></br></br><br />
The curve of I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub> contradicted our common sense severely. First, I<sub>m</sub>>C<sub>i</sub>>R<sub>i</sub> is expected to be tenable all the time, which precludes the intersects among the three curves; Second, there is no mechanism in this system that could decrease their concentration, and all of them are increasing function; Third and most serious, never will them be negative, as they represent the concentration of real substances.<br />
<br />
Then we adjusted the parameters slightly for several times. To eliminate those absurd curves, we reconsidered some assumptions.<br />
Here we listed another representative group of parament values and relative graph:</div></br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/7/7d/Suicide3.png/800px-Suicide3.png"></br></br><br />
In this group, we gave up one former assumption and set k<sub>2</sub> equal to k<sub>9</sub>. We also gave positive values to I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub>, which were considered to be zero at first. And by groups of stimulations we realized the value of k<sub>2</sub> does matter, as the derivative of C<sub>f</sub> only increased slightly as k<sub>2</sub> lowers, and the positive values failed to avoid the weird phenomenon in the latter three curves.<br />
We also found that however we adjusted the primary value of I<sub>f</sub> and other parameters, If dropped into approximately zero extremely rapidly at the initial stage and remained balanced, which might account for why the derivatives of the latter curves were abnormally negative. Thus we modified another assumption and increased k<sub>7</sub>. Here is another group of values and corresponding graph:</br></br><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/1/15/Suicide4.png/800px-Suicide4.png" style="width:600; height:400;"></br></br><br />
Although the derivative of Im is not seriously positive constantly, the three latter curves seemed much more reasonable. Hence, we extrapolated although SdpI and SdpR share the same promoter, the expression of SdpI must much faster than SdpR to ensure successful “suicide.” Additionally, the increase of k<sub>7</sub> also represses SdpC, and hence the copy number of SdpC must be larger.<br />
We kept all other parameters constant and gradually augmented k<sub>0</sub>. The larger k<sub>0</sub>, the more perfect the curve seemed, and here are the values table and graph where k<sub>0</sub> equals 400, 80 times larger than k<sub>4</sub>.</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>400</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/3/33/Suicide5.png/800px-Suicide5.png"></br></br><br />
Take the curve of C<sub>f</sub> and R<sub>f</sub> separately, the curves seemed more perfect:</br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/9/9f/Suicide6.png"></br></br><br />
<br />
In wild bacteria who are unable to produced SdpC, naturally k<sub>0</sub> equals zero. We expected C<sub>f</sub> would decreased gradually and finally approximate zero, and here are the corresponding table and graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>30</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/1/19/Suicide7.png"></br></br><br />
Wired but not surprising, there were intersects among the latter three curve, and C<sub>f</sub> decreases continually when it is negative. We continued to try groups of these parameters, and this is the best one where we increased the primary concentration of SdpC and the normal expression rate of SdpI.<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/8/8e/Suicide8.png"></br></br><br />
The curve of C<sub>f</sub> and R<sub>f</sub> alone:<br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/e/eb/Suicide9.png"></br></br><br />
In spite of minimal abnormal phenomenon (C<sub>f</sub> was negative in later stage), this graph roughly testified that in wild bacterial the concentration of float SdpC will drop to nearly zero quickly.<br />
In sum, the ODE model of singular cells indicate following results:</br><br />
<ol><br />
<li>The character of free SdpC is most affected by k<sub>0</sub>, if the copy number of SdpC is large enough, it is theoretically reasonable to commit suicide;</li><br />
<li>The influence of the value of Imax and k<sub>2</sub> is much limited;</li><br />
<li>The amount of free SdpI is always near zero;</li><br />
<li>SdpC will not increase limitlessly however we transform parameters;</li><br />
<li>To ensure the success of suicide, it is required k<sub>0</sub>>>k<sub>4</sub>>>k<sub>7</sub>;</li><br />
The last conclusion was our biggest windfall, and we have verified the validity of this suicide mechanism in math. On the one hand, if further experiments proven #4 engineered bacteria will kill both siblings and themselves, it is highly like that the expression rate SdpI is much larger than SdpR even if they share the same promoter; on the other hand, if #4 engineered bacteria are not able to commit suicide, we can try to boost the expression of SdpI to adjust the bacteria.<br />
</ol><br />
<br />
<br />
<h1>Discussion on colonies</h1><br />
<br />
In reality, the engineered bacteria aims at killing its siblings instead of itself, and at first almost all toxin SdpC will be secreted outside the bacteria. Assume the diffusion of toxin among cells comply with diffusion law, that is, the diffusion rate is proportionate with the gradient of concentration. Further assume the death concentration of SdpC is same to all bacteria expect those who contain this locus, the average life expectancy is bacteria will hinge on the rate and distribution of engineered bacteria, and the distribution of life expectancy of bacteria is similar to that of average free path of gas molecules.<br />
As long the coefficient of diffusion is large enough, any engineered bacterias, no matter how few, is adequate to devastate the whole colony. Alike to the average free path of thin gas, the average suicide time of the whole reporter system is inversely proportional with the square root of the rate of engineer bacteria containing this locus.<br />
<br />
<br />
<h1>References</h1><br />
Parallel pathways of repression and antirepression governing the transition to stationary phase in Bacillus subtilis<br />
AV Banse, A Chastanet, L Rahn-Lee…,PNAS ,2008 </br><br />
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<h1>Introduction</h1><br />
To be more user-friendly, 4# circuit contains a reporting system. After melting in water, the spores germinate and express blue pigment protein to report the best using time.</br><br />
Meanwhile, 4# circuit could also ensure biosafety. Because other circuit do not have self-killing device, 4# engineering bacterial should kill all the bacterial after using.</br><br />
<br />
</div><br />
<br />
<div><br />
<h1>Designing of the suicide system</h1><br />
We design a circuit of killing switch based on its endogenous genetic system.</br><br />
In B.subtilis, when it comes to the stationary phase, the environmental pressure increases and nutrition becomes limited, so B begin to produce spores. Now the community will be divided into two different parts. One of them are trying to kill others to get enough nutrient , delaying the production of spores and achieving a competitive advantage. Killing is mediated by the exported toxic protein SdpC. SdpI will appear on the membrane surface to avoid itself from being damaged. SdpI could bind free SdpC and autopressor SdpR, to remove SdpR’s inhibition against I and R, to produce more SdpI to offset SdpC, finally guaranteeing the subgroup alive, thereby delaying the spores production.</br><br />
<img src="https://static.igem.org/mediawiki/2013/2/2b/Reporter_3.png" width="500" height="350"><br />
<p>We transfer SdpC which is fused by promoter SdpI/R into high copy plasmids in order to damage the balance of the system, thereby killing whole colony. When SdpC appears, SdpI on the membrane will bind free SdpC and adsorb SdpR to cease its inhibition against SdpI P/R, trying to produce more SdpI. At the same time, it will activate the promoter SdpR/I in our circuits and generate more SdpC.The system would fall into an infinite loop, and according to our modeling ,the amount of SdpC increases beyond the ability of SdpI.Thus,the cells with protection mechanism will crack and die because of too much SdpC. All above forms the killing device.<br />
We Also designed a test circuit,which contains promotor grac and sdpABC only,aiming to determine the ability of SdpC.</p><br />
<br />
<div></br></br><br />
There are both positive and negative feedback loops in this process. On the one hand, SdpI is unable to sequestrate the autorepressor, SdpR, until it captures the toxin, SdpC. The accumulation of SdpC will thus facilitate SdpI to capture more SdpR and thereby relieve the repression of SdpR, stimulating the expression of itself. This is the positive feedback loop which leads to the increasing accumulation of SdpC and finally the death of the bacteria. On the other hand, the removal of SdpR also enhance the expression of SdpI and accelerate the sequestration of SdpC, which forms a negative feedback loop whose effects contradict the positive feedback loop. However, since the copy number of SdpC is much higher, it is believed that the positive loop is strong enough to outweigh the negative one, which guarantees this mechanism will finally leads to collapse instead of equilibrium.<br />
</div><br />
<div><br />
<h1>The ODE model of singular cells</h1><br />
There is no denying fact that the essential goal of engineered bacterias who carry this so called “suicide” locus itself is to kill their siblings rather than themselves to ensure the survival of themselves. Surly they can kill their siblings, but can they finally eliminate themselves, as we expects? The trivial experiment protocol and huge uncertainty had put off our experiment, and as expected, we failed to achieve the construction of complete reporter system in our laboratory. Fortunately, we could resort to mathematical models to verify the validity of this locus theoretically. <br />
There are six independent variables in individual cells, and the theoretically if the initial conditions are fixed, all of them will be the univariate functions of time. The following table illustrates the mark and meaning of each variable:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub></td><br />
<td >Mole number of free SdpI in cytoplasm</a>.</td><br />
</tr><br />
<tr><br />
<td >I<sub>m</sub> </td><br />
<td >Mole number of SdpI in the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >C<sub>f</sub></td><br />
<td >Mole number of free SdpC in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >C<sub>i</sub></td><br />
<td >Mole number of SdpC captured by SdpI.</td><br />
</tr><br />
<tr><br />
<td >R<sub>f</sub></td><br />
<td >Mole number of free SdpR in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >R<sub>i</sub> </td><br />
<td >Mole number of SdpR captured by SdpI</td><br />
</tr><br />
</table><br />
</br></br><br />
<div>To construct reasonable ordinary differential equation (ODE) model to describe and predict the operation of the suicide system, we followed the law of mass action, one basic law of chemistry and biology.</br><br />
Taken as a statement about kinetics, the law states that the rate of an elementary reaction (a reaction that proceeds through only one transition state, which is one mechanistic step) is proportional to the product of the concentrations of the participating molecules. In modern chemistry this is derived using statistical mechanics. Despite the complicated chemical reactions involved in the process of transcription and translation, it is common and logically sound to view the expression of one particular gene as an elementary reaction and assume the repression effects of the protein itself encodes and the repressor are both linear.</br><br />
According to the law of mass action, we got six independent differential equation of the variables:</br><br />
<img src="https://static.igem.org/mediawiki/2013/e/e8/For1%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/2/23/For2%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/4/48/For3%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/7/71/For4%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/3/31/For5%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/c/cb/For6%281%29.png"></br></br></br><br />
The following table explain the constants in the above ODE groups:</br></br></div><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub> </td><br />
<td >The maximal number of SdpI than can be fixed on the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >k<sub>0</sub></td><br />
<td >Constant describes the normal expression rate of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>1</sub> </td><br />
<td >Constant describes the self-repression effects of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>2</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpC.</a></td><br />
</tr><br />
<tr><br />
<td >k<sub>3</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>4</sub> </td><br />
<td >Constant describes the normal expression rate of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>5</sub> </td><br />
<td >Constant describes the self-repression effects of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>6</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>7</sub> </td><br />
<td >Constant describes the normal expression rate of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>8</sub> </td><br />
<td >Constant describes the self-repression effects of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>9</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>10</sub> </td><br />
<td >Constant describes the rate of SdpI binding to the cell membrane</td><br />
</tr><br />
</table><br />
<br />
<br />
<br />
<br />
</div><br />
<div class="clear"></div><br />
<div><br />
<h2>Discussions on the constants</h2><br />
All the constants given above is steady and theoretically measurable when all the conditions are constant. For example, we could measure k<sub>0</sub> by constructing a new engineered bacteria, which contains the gene encoding SdpC and marker gene alone and observing the influence of the concentration of SdpC on its expression. Yet any modification on genome is notoriously time-consuming, which inhibited us from measuring them in person. We also looked up oceans of papers to confer their approximate ranges, but almost all papers are too fragmental to afford any valid information. Therefore, we decided to assume all these constant according to our limited information and make a qualitative analysis instead of quantifiable analysis. All units and dimensions were temporarily ignored. In other words, our model aims at justifying the validity of this suicide mechanism rather than predicting the exact time or any other parameters of the system.<br />
<br />
Despite the fact that we have hardly any accurate data on these constants, there are some limitations that we extrapolated from known information before we further explore this model:</br><br />
<ol><br />
<li>k<sub>0</sub>>>k<sub>4</sub>≈k<sub>7</sub>: k<sub>0</sub>,k<sub>4</sub> and k<sub>7</sub> represent the normal expression rate of SdpC, SdpR and SdpC separately, and the copy number of SdpC is much larger than that of SdpR and SdpI, whereas the value of the latter two is approximately equal;</li><br />
<li>k<sub>2</sub>>>k<sub>9</sub>: the existence of free SdpR represses the expression of both SdpI and SdpC, and similarly, since the copy number of SdpC is much higher, we expected the repression effect was stronger accordingly;</li><br />
<li>k<sub>10</sub>>>k<sub>3</sub>,k<sub>6</sub>:it is hard to predict the value of k<sub>3</sub> and k<sub>8</sub>, yet we suppose both of them is much smaller than k<sub>10</sub> because SdpI is a kind of membrane protein inherently, and rarely exists as free protein</li><br />
<li>The primary values of all the six variables are very small or strictly zero. We expect it as the most logical initial status. If the primary value of any variable is relatively large, the suicide mechanism may not run normally</li><br />
</ol><br />
</br></br><br />
<h2>Stimulation and discussion</h2><br />
<br />
Simple and rough as the above model is, it does theoretically sound. To test the validity of this model, we first tried to get analytic solution of the ODE set. If this analytic solution exists, we could further investigate the interaction among those variables, and draw some phase planes to get accurate and mathematically perfect description of this model. Unfortunately but expectedly, the existence of analytic solution was negated by MATLAB, and we had to assume groups of values for these constants in advance and analyze the arithmetic solutions instead. These arithmetic solutions not only justified this mechanism is effective enough to commit cell suicide but also indicated some unexpected, or even weird results that beyond our wildest imagination. There are two possibility account for the unexpected results: our model is too rough to include some assignable factor; or there are some implicit but objective limitation inside model, which may be substantiate by later experiments or papers.</br><br />
When we explored the arithmetic solutions of this ODE set, we received nearly one hundred warnings from MATLAB and for many times our most powerful computer ran out of its 8GB memory, but sometimes we can receive the solution within seconds. We had adjusted our parameters for several times before we got our first solution. Here is the values of parameters for this group, and the graph of arithmetic solutions is also given:</br><br />
<div style="margin-top:20px;"><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>50</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table><br />
</div></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/2/26/Suicide1.png"></br></br><br />
At the first glance this graph seemed fine. Initially the concentration of SdpC decreased slightly due to the capturing of SdpI and the repression of float SdpR, but gradually the positive feedback loop works, and C<sub>f</sub> increases rapidly. But when we turned our attention to the curves of other parameters, things seemed not so perfect:</br><br />
<div><img class="linegraph" src="https://static.igem.org/mediawiki/2013/c/cf/Suicide2.png"></br></br><br />
The curve of I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub> contradicted our common sense severely. First, I<sub>m</sub>>C<sub>i</sub>>R<sub>i</sub> is expected to be tenable all the time, which precludes the intersects among the three curves; Second, there is no mechanism in this system that could decrease their concentration, and all of them are increasing function; Third and most serious, never will them be negative, as they represent the concentration of real substances.<br />
<br />
Then we adjusted the parameters slightly for several times. To eliminate those absurd curves, we reconsidered some assumptions.<br />
Here we listed another representative group of parament values and relative graph:</div></br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/7/7d/Suicide3.png/800px-Suicide3.png"></br></br><br />
In this group, we gave up one former assumption and set k<sub>2</sub> equal to k<sub>9</sub>. We also gave positive values to I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub>, which were considered to be zero at first. And by groups of stimulations we realized the value of k<sub>2</sub> does matter, as the derivative of C<sub>f</sub> only increased slightly as k<sub>2</sub> lowers, and the positive values failed to avoid the weird phenomenon in the latter three curves.<br />
We also found that however we adjusted the primary value of I<sub>f</sub> and other parameters, If dropped into approximately zero extremely rapidly at the initial stage and remained balanced, which might account for why the derivatives of the latter curves were abnormally negative. Thus we modified another assumption and increased k<sub>7</sub>. Here is another group of values and corresponding graph:</br></br><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/1/15/Suicide4.png/800px-Suicide4.png" style="width:600; height:400;"></br></br><br />
Although the derivative of Im is not seriously positive constantly, the three latter curves seemed much more reasonable. Hence, we extrapolated although SdpI and SdpR share the same promoter, the expression of SdpI must much faster than SdpR to ensure successful “suicide.” Additionally, the increase of k<sub>7</sub> also represses SdpC, and hence the copy number of SdpC must be larger.<br />
We kept all other parameters constant and gradually augmented k<sub>0</sub>. The larger k<sub>0</sub>, the more perfect the curve seemed, and here are the values table and graph where k<sub>0</sub> equals 400, 80 times larger than k<sub>4</sub>.</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>400</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/3/33/Suicide5.png/800px-Suicide5.png"></br></br><br />
Take the curve of C<sub>f</sub> and R<sub>f</sub> separately, the curves seemed more perfect:</br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/9/9f/Suicide6.png"></br></br><br />
<br />
In wild bacteria who are unable to produced SdpC, naturally k<sub>0</sub> equals zero. We expected C<sub>f</sub> would decreased gradually and finally approximate zero, and here are the corresponding table and graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>30</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/1/19/Suicide7.png"></br></br><br />
Wired but not surprising, there were intersects among the latter three curve, and C<sub>f</sub> decreases continually when it is negative. We continued to try groups of these parameters, and this is the best one where we increased the primary concentration of SdpC and the normal expression rate of SdpI.<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/8/8e/Suicide8.png"></br></br><br />
The curve of C<sub>f</sub> and R<sub>f</sub> alone:<br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/e/eb/Suicide9.png"></br></br><br />
In spite of minimal abnormal phenomenon (C<sub>f</sub> was negative in later stage), this graph roughly testified that in wild bacterial the concentration of float SdpC will drop to nearly zero quickly.<br />
In sum, the ODE model of singular cells indicate following results:</br><br />
<ol><br />
<li>The character of free SdpC is most affected by k<sub>0</sub>, if the copy number of SdpC is large enough, it is theoretically reasonable to commit suicide;</li><br />
<li>The influence of the value of Imax and k<sub>2</sub> is much limited;</li><br />
<li>The amount of free SdpI is always near zero;</li><br />
<li>SdpC will not increase limitlessly however we transform parameters;</li><br />
<li>To ensure the success of suicide, it is required k<sub>0</sub>>>k<sub>4</sub>>>k<sub>7</sub>;</li><br />
The last conclusion was our biggest windfall, and we have verified the validity of this suicide mechanism in math. On the one hand, if further experiments proven #4 engineered bacteria will kill both siblings and themselves, it is highly like that the expression rate SdpI is much larger than SdpR even if they share the same promoter; on the other hand, if #4 engineered bacteria are not able to commit suicide, we can try to boost the expression of SdpI to adjust the bacteria.<br />
<br />
<br />
<h1>Discussion on colonies</h1><br />
<br />
In reality, the engineered bacteria aims at killing its siblings instead of itself, and at first almost all toxin SdpC will be secreted outside the bacteria. Assume the diffusion of toxin among cells comply with diffusion law, that is, the diffusion rate is proportionate with the gradient of concentration. Further assume the death concentration of SdpC is same to all bacteria expect those who contain this locus, the average life expectancy is bacteria will hinge on the rate and distribution of engineered bacteria, and the distribution of life expectancy of bacteria is similar to that of average free path of gas molecules.<br />
As long the coefficient of diffusion is large enough, any engineered bacterias, no matter how few, is adequate to devastate the whole colony. Alike to the average free path of thin gas, the average suicide time of the whole reporter system is inversely proportional with the square root of the rate of engineer bacteria containing this locus.<br />
<br />
<br />
<h1>References</h1><br />
Parallel pathways of repression and antirepression governing the transition to stationary phase in Bacillus subtilis<br />
AV Banse, A Chastanet, L Rahn-Lee…,PNAS ,2008 </br><br />
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<h1>Introduction</h1><br />
To be more user-friendly, 4# circuit contains a reporting system. After melting in water, the spores germinate and express blue pigment protein to report the best using time.</br><br />
Meanwhile, 4# circuit could also ensure biosafety. Because other circuit do not have self-killing device, 4# engineering bacterial should kill all the bacterial after using.</br><br />
<br />
</div><br />
<br />
<div><br />
<h1>Designing of the suicide system</h1><br />
We design a circuit of killing switch based on its endogenous genetic system.</br><br />
In B.subtilis, when it comes to the stationary phase, the environmental pressure increases and nutrition becomes limited, so B begin to produce spores. Now the community will be divided into two different parts. One of them are trying to kill others to get enough nutrient , delaying the production of spores and achieving a competitive advantage. Killing is mediated by the exported toxic protein SdpC. SdpI will appear on the membrane surface to avoid itself from being damaged. SdpI could bind free SdpC and autopressor SdpR, to remove SdpR’s inhibition against I and R, to produce more SdpI to offset SdpC, finally guaranteeing the subgroup alive, thereby delaying the spores production.</br><br />
<img src="https://static.igem.org/mediawiki/2013/2/2b/Reporter_3.png" width="500" height="350"><br />
<p>We transfer SdpC which is fused by promoter SdpI/R into high copy plasmids in order to damage the balance of the system, thereby killing whole colony. When SdpC appears, SdpI on the membrane will bind free SdpC and adsorb SdpR to cease its inhibition against SdpI P/R, trying to produce more SdpI. At the same time, it will activate the promoter SdpR/I in our circuits and generate more SdpC.The system would fall into an infinite loop, and according to our modeling ,the amount of SdpC increases beyond the ability of SdpI.Thus,the cells with protection mechanism will crack and die because of too much SdpC. All above forms the killing device.<br />
We Also designed a test circuit,which contains promotor grac and sdpABC only,aiming to determine the ability of SdpC.</p><br />
<br />
<div></br></br><br />
There are both positive and negative feedback loops in this process. On the one hand, SdpI is unable to sequestrate the autorepressor, SdpR, until it captures the toxin, SdpC. The accumulation of SdpC will thus facilitate SdpI to capture more SdpR and thereby relieve the repression of SdpR, stimulating the expression of itself. This is the positive feedback loop which leads to the increasing accumulation of SdpC and finally the death of the bacteria. On the other hand, the removal of SdpR also enhance the expression of SdpI and accelerate the sequestration of SdpC, which forms a negative feedback loop whose effects contradict the positive feedback loop. However, since the copy number of SdpC is much higher, it is believed that the positive loop is strong enough to outweigh the negative one, which guarantees this mechanism will finally leads to collapse instead of equilibrium.<br />
</div><br />
<div><br />
<h1>The ODE model of singular cells</h1><br />
There is no denying fact that the essential goal of engineered bacterias who carry this so called “suicide” locus itself is to kill their siblings rather than themselves to ensure the survival of themselves. Surly they can kill their siblings, but can they finally eliminate themselves, as we expects? The trivial experiment protocol and huge uncertainty had put off our experiment, and as expected, we failed to achieve the construction of complete reporter system in our laboratory. Fortunately, we could resort to mathematical models to verify the validity of this locus theoretically. <br />
There are six independent variables in individual cells, and the theoretically if the initial conditions are fixed, all of them will be the univariate functions of time. The following table illustrates the mark and meaning of each variable:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub></td><br />
<td >Mole number of free SdpI in cytoplasm</a>.</td><br />
</tr><br />
<tr><br />
<td >I<sub>m</sub> </td><br />
<td >Mole number of SdpI in the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >C<sub>f</sub></td><br />
<td >Mole number of free SdpC in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >C<sub>i</sub></td><br />
<td >Mole number of SdpC captured by SdpI.</td><br />
</tr><br />
<tr><br />
<td >R<sub>f</sub></td><br />
<td >Mole number of free SdpR in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >R<sub>i</sub> </td><br />
<td >Mole number of SdpR captured by SdpI</td><br />
</tr><br />
</table><br />
</br></br><br />
<div>To construct reasonable ordinary differential equation (ODE) model to describe and predict the operation of the suicide system, we followed the law of mass action, one basic law of chemistry and biology.</br><br />
Taken as a statement about kinetics, the law states that the rate of an elementary reaction (a reaction that proceeds through only one transition state, which is one mechanistic step) is proportional to the product of the concentrations of the participating molecules. In modern chemistry this is derived using statistical mechanics. Despite the complicated chemical reactions involved in the process of transcription and translation, it is common and logically sound to view the expression of one particular gene as an elementary reaction and assume the repression effects of the protein itself encodes and the repressor are both linear.</br><br />
According to the law of mass action, we got six independent differential equation of the variables:</br><br />
<img src="https://static.igem.org/mediawiki/2013/e/e8/For1%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/2/23/For2%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/4/48/For3%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/7/71/For4%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/3/31/For5%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/c/cb/For6%281%29.png"></br></br></br><br />
The following table explain the constants in the above ODE groups:</br></br></div><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub> </td><br />
<td >The maximal number of SdpI than can be fixed on the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >k<sub>0</sub></td><br />
<td >Constant describes the normal expression rate of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>1</sub> </td><br />
<td >Constant describes the self-repression effects of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>2</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpC.</a></td><br />
</tr><br />
<tr><br />
<td >k<sub>3</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>4</sub> </td><br />
<td >Constant describes the normal expression rate of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>5</sub> </td><br />
<td >Constant describes the self-repression effects of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>6</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>7</sub> </td><br />
<td >Constant describes the normal expression rate of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>8</sub> </td><br />
<td >Constant describes the self-repression effects of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>9</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>10</sub> </td><br />
<td >Constant describes the rate of SdpI binding to the cell membrane</td><br />
</tr><br />
</table><br />
<br />
<br />
<br />
<br />
</div><br />
<div><br />
<h2>Discussions on the constants</h2><br />
All the constants given above is steady and theoretically measurable when all the conditions are constant. For example, we could measure k<sub>0</sub> by constructing a new engineered bacteria, which contains the gene encoding SdpC and marker gene alone and observing the influence of the concentration of SdpC on its expression. Yet any modification on genome is notoriously time-consuming, which inhibited us from measuring them in person. We also looked up oceans of papers to confer their approximate ranges, but almost all papers are too fragmental to afford any valid information. Therefore, we decided to assume all these constant according to our limited information and make a qualitative analysis instead of quantifiable analysis. All units and dimensions were temporarily ignored. In other words, our model aims at justifying the validity of this suicide mechanism rather than predicting the exact time or any other parameters of the system.<br />
<br />
Despite the fact that we have hardly any accurate data on these constants, there are some limitations that we extrapolated from known information before we further explore this model:</br><br />
<ol><br />
<li>k<sub>0</sub>>>k<sub>4</sub>≈k<sub>7</sub>: k<sub>0</sub>,k<sub>4</sub> and k<sub>7</sub> represent the normal expression rate of SdpC, SdpR and SdpC separately, and the copy number of SdpC is much larger than that of SdpR and SdpI, whereas the value of the latter two is approximately equal;</li><br />
<li>k<sub>2</sub>>>k<sub>9</sub>: the existence of free SdpR represses the expression of both SdpI and SdpC, and similarly, since the copy number of SdpC is much higher, we expected the repression effect was stronger accordingly;</li><br />
<li>k<sub>10</sub>>>k<sub>3</sub>,k<sub>6</sub>:it is hard to predict the value of k<sub>3</sub> and k<sub>8</sub>, yet we suppose both of them is much smaller than k<sub>10</sub> because SdpI is a kind of membrane protein inherently, and rarely exists as free protein</li><br />
<li>The primary values of all the six variables are very small or strictly zero. We expect it as the most logical initial status. If the primary value of any variable is relatively large, the suicide mechanism may not run normally</li><br />
</ol><br />
</br></br><br />
<h2>Stimulation and discussion</h2><br />
<br />
Simple and rough as the above model is, it does theoretically sound. To test the validity of this model, we first tried to get analytic solution of the ODE set. If this analytic solution exists, we could further investigate the interaction among those variables, and draw some phase planes to get accurate and mathematically perfect description of this model. Unfortunately but expectedly, the existence of analytic solution was negated by MATLAB, and we had to assume groups of values for these constants in advance and analyze the arithmetic solutions instead. These arithmetic solutions not only justified this mechanism is effective enough to commit cell suicide but also indicated some unexpected, or even weird results that beyond our wildest imagination. There are two possibility account for the unexpected results: our model is too rough to include some assignable factor; or there are some implicit but objective limitation inside model, which may be substantiate by later experiments or papers.</br><br />
When we explored the arithmetic solutions of this ODE set, we received nearly one hundred warnings from MATLAB and for many times our most powerful computer ran out of its 8GB memory, but sometimes we can receive the solution within seconds. We had adjusted our parameters for several times before we got our first solution. Here is the values of parameters for this group, and the graph of arithmetic solutions is also given:</br><br />
<div style="margin-top:20px;"><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>50</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table><br />
</div></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/2/26/Suicide1.png"></br></br><br />
At the first glance this graph seemed fine. Initially the concentration of SdpC decreased slightly due to the capturing of SdpI and the repression of float SdpR, but gradually the positive feedback loop works, and C<sub>f</sub> increases rapidly. But when we turned our attention to the curves of other parameters, things seemed not so perfect:</br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/c/cf/Suicide2.png"></br></br><br />
The curve of I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub> contradicted our common sense severely. First, I<sub>m</sub>>C<sub>i</sub>>R<sub>i</sub> is expected to be tenable all the time, which precludes the intersects among the three curves; Second, there is no mechanism in this system that could decrease their concentration, and all of them are increasing function; Third and most serious, never will them be negative, as they represent the concentration of real substances.<br />
<br />
Then we adjusted the parameters slightly for several times. To eliminate those absurd curves, we reconsidered some assumptions.<br />
Here we listed another representative group of parament values and relative graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/7/7d/Suicide3.png/800px-Suicide3.png"></br></br><br />
In this group, we gave up one former assumption and set k<sub>2</sub> equal to k<sub>9</sub>. We also gave positive values to I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub>, which were considered to be zero at first. And by groups of stimulations we realized the value of k<sub>2</sub> does matter, as the derivative of C<sub>f</sub> only increased slightly as k<sub>2</sub> lowers, and the positive values failed to avoid the weird phenomenon in the latter three curves.<br />
We also found that however we adjusted the primary value of I<sub>f</sub> and other parameters, If dropped into approximately zero extremely rapidly at the initial stage and remained balanced, which might account for why the derivatives of the latter curves were abnormally negative. Thus we modified another assumption and increased k<sub>7</sub>. Here is another group of values and corresponding graph:</br></br><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/1/15/Suicide4.png/800px-Suicide4.png" style="width:600; height:400;"></br></br><br />
Although the derivative of Im is not seriously positive constantly, the three latter curves seemed much more reasonable. Hence, we extrapolated although SdpI and SdpR share the same promoter, the expression of SdpI must much faster than SdpR to ensure successful “suicide.” Additionally, the increase of k<sub>7</sub> also represses SdpC, and hence the copy number of SdpC must be larger.<br />
We kept all other parameters constant and gradually augmented k<sub>0</sub>. The larger k<sub>0</sub>, the more perfect the curve seemed, and here are the values table and graph where k<sub>0</sub> equals 400, 80 times larger than k<sub>4</sub>.</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>400</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/3/33/Suicide5.png/800px-Suicide5.png"></br></br><br />
Take the curve of C<sub>f</sub> and R<sub>f</sub> separately, the curves seemed more perfect:</br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/9/9f/Suicide6.png"></br></br><br />
<br />
In wild bacteria who are unable to produced SdpC, naturally k<sub>0</sub> equals zero. We expected C<sub>f</sub> would decreased gradually and finally approximate zero, and here are the corresponding table and graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>30</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/1/19/Suicide7.png"></br></br><br />
Wired but not surprising, there were intersects among the latter three curve, and C<sub>f</sub> decreases continually when it is negative. We continued to try groups of these parameters, and this is the best one where we increased the primary concentration of SdpC and the normal expression rate of SdpI.<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/8/8e/Suicide8.png"></br></br><br />
The curve of C<sub>f</sub> and R<sub>f</sub> alone:<br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/e/eb/Suicide9.png"></br></br><br />
In spite of minimal abnormal phenomenon (C<sub>f</sub> was negative in later stage), this graph roughly testified that in wild bacterial the concentration of float SdpC will drop to nearly zero quickly.<br />
In sum, the ODE model of singular cells indicate following results:</br><br />
<ol><br />
<li>The character of free SdpC is most affected by k<sub>0</sub>, if the copy number of SdpC is large enough, it is theoretically reasonable to commit suicide;</li><br />
<li>The influence of the value of Imax and k<sub>2</sub> is much limited;</li><br />
<li>The amount of free SdpI is always near zero;</li><br />
<li>SdpC will not increase limitlessly however we transform parameters;</li><br />
<li>To ensure the success of suicide, it is required k<sub>0</sub>>>k<sub>4</sub>>>k<sub>7</sub>;</li><br />
The last conclusion was our biggest windfall, and we have verified the validity of this suicide mechanism in math. On the one hand, if further experiments proven #4 engineered bacteria will kill both siblings and themselves, it is highly like that the expression rate SdpI is much larger than SdpR even if they share the same promoter; on the other hand, if #4 engineered bacteria are not able to commit suicide, we can try to boost the expression of SdpI to adjust the bacteria.<br />
<br />
<br />
<h1>Discussion on colonies</h1><br />
<br />
In reality, the engineered bacteria aims at killing its siblings instead of itself, and at first almost all toxin SdpC will be secreted outside the bacteria. Assume the diffusion of toxin among cells comply with diffusion law, that is, the diffusion rate is proportionate with the gradient of concentration. Further assume the death concentration of SdpC is same to all bacteria expect those who contain this locus, the average life expectancy is bacteria will hinge on the rate and distribution of engineered bacteria, and the distribution of life expectancy of bacteria is similar to that of average free path of gas molecules.<br />
As long the coefficient of diffusion is large enough, any engineered bacterias, no matter how few, is adequate to devastate the whole colony. Alike to the average free path of thin gas, the average suicide time of the whole reporter system is inversely proportional with the square root of the rate of engineer bacteria containing this locus.<br />
<br />
<br />
<h1>References</h1><br />
Parallel pathways of repression and antirepression governing the transition to stationary phase in Bacillus subtilis<br />
AV Banse, A Chastanet, L Rahn-Lee…,PNAS ,2008 </br><br />
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<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a>&gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></div></div><br />
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<h1>Introduction</h1><br />
To be more user-friendly, 4# circuit contains a reporting system. After melting in water, the spores germinate and express blue pigment protein to report the best using time.</br><br />
Meanwhile, 4# circuit could also ensure biosafety. Because other circuit do not have self-killing device, 4# engineering bacterial should kill all the bacterial after using.</br><br />
<br />
</div><br />
<br />
<div><br />
<h1>Designing of the suicide system</h1><br />
We design a circuit of killing switch based on its endogenous genetic system.</br><br />
In B.subtilis, when it comes to the stationary phase, the environmental pressure increases and nutrition becomes limited, so B begin to produce spores. Now the community will be divided into two different parts. One of them are trying to kill others to get enough nutrient , delaying the production of spores and achieving a competitive advantage. Killing is mediated by the exported toxic protein SdpC. SdpI will appear on the membrane surface to avoid itself from being damaged. SdpI could bind free SdpC and autopressor SdpR, to remove SdpR’s inhibition against I and R, to produce more SdpI to offset SdpC, finally guaranteeing the subgroup alive, thereby delaying the spores production.</br><br />
<img src="https://static.igem.org/mediawiki/2013/2/2b/Reporter_3.png" width="500" height="350"><br />
<p>We transfer SdpC which is fused by promoter SdpI/R into high copy plasmids in order to damage the balance of the system, thereby killing whole colony. When SdpC appears, SdpI on the membrane will bind free SdpC and adsorb SdpR to cease its inhibition against SdpI P/R, trying to produce more SdpI. At the same time, it will activate the promoter SdpR/I in our circuits and generate more SdpC.The system would fall into an infinite loop, and according to our modeling ,the amount of SdpC increases beyond the ability of SdpI.Thus,the cells with protection mechanism will crack and die because of too much SdpC. All above forms the killing device.<br />
We Also designed a test circuit,which contains promotor grac and sdpABC only,aiming to determine the ability of SdpC.</p><br />
<br />
<div></br></br><br />
There are both positive and negative feedback loops in this process. On the one hand, SdpI is unable to sequestrate the autorepressor, SdpR, until it captures the toxin, SdpC. The accumulation of SdpC will thus facilitate SdpI to capture more SdpR and thereby relieve the repression of SdpR, stimulating the expression of itself. This is the positive feedback loop which leads to the increasing accumulation of SdpC and finally the death of the bacteria. On the other hand, the removal of SdpR also enhance the expression of SdpI and accelerate the sequestration of SdpC, which forms a negative feedback loop whose effects contradict the positive feedback loop. However, since the copy number of SdpC is much higher, it is believed that the positive loop is strong enough to outweigh the negative one, which guarantees this mechanism will finally leads to collapse instead of equilibrium.<br />
</div><br />
<div><br />
<h1>The ODE model of singular cells</h1><br />
There is no denying fact that the essential goal of engineered bacterias who carry this so called “suicide” locus itself is to kill their siblings rather than themselves to ensure the survival of themselves. Surly they can kill their siblings, but can they finally eliminate themselves, as we expects? The trivial experiment protocol and huge uncertainty had put off our experiment, and as expected, we failed to achieve the construction of complete reporter system in our laboratory. Fortunately, we could resort to mathematical models to verify the validity of this locus theoretically. <br />
There are six independent variables in individual cells, and the theoretically if the initial conditions are fixed, all of them will be the univariate functions of time. The following table illustrates the mark and meaning of each variable:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub></td><br />
<td >Mole number of free SdpI in cytoplasm</a>.</td><br />
</tr><br />
<tr><br />
<td >I<sub>m</sub> </td><br />
<td >Mole number of SdpI in the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >C<sub>f</sub></td><br />
<td >Mole number of free SdpC in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >C<sub>i</sub></td><br />
<td >Mole number of SdpC captured by SdpI.</td><br />
</tr><br />
<tr><br />
<td >R<sub>f</sub></td><br />
<td >Mole number of free SdpR in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >R<sub>i</sub> </td><br />
<td >Mole number of SdpR captured by SdpI</td><br />
</tr><br />
</table><br />
</br></br><br />
<div>To construct reasonable ordinary differential equation (ODE) model to describe and predict the operation of the suicide system, we followed the law of mass action, one basic law of chemistry and biology.</br><br />
Taken as a statement about kinetics, the law states that the rate of an elementary reaction (a reaction that proceeds through only one transition state, which is one mechanistic step) is proportional to the product of the concentrations of the participating molecules. In modern chemistry this is derived using statistical mechanics. Despite the complicated chemical reactions involved in the process of transcription and translation, it is common and logically sound to view the expression of one particular gene as an elementary reaction and assume the repression effects of the protein itself encodes and the repressor are both linear.</br><br />
According to the law of mass action, we got six independent differential equation of the variables:</br><br />
<img src="https://static.igem.org/mediawiki/2013/e/e8/For1%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/2/23/For2%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/4/48/For3%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/7/71/For4%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/3/31/For5%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/c/cb/For6%281%29.png"></br></br></br><br />
The following table explain the constants in the above ODE groups:</br></br></div><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub> </td><br />
<td >The maximal number of SdpI than can be fixed on the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >k<sub>0</sub></td><br />
<td >Constant describes the normal expression rate of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>1</sub> </td><br />
<td >Constant describes the self-repression effects of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>2</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpC.</a></td><br />
</tr><br />
<tr><br />
<td >k<sub>3</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>4</sub> </td><br />
<td >Constant describes the normal expression rate of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>5</sub> </td><br />
<td >Constant describes the self-repression effects of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>6</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>7</sub> </td><br />
<td >Constant describes the normal expression rate of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>8</sub> </td><br />
<td >Constant describes the self-repression effects of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>9</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>10</sub> </td><br />
<td >Constant describes the rate of SdpI binding to the cell membrane</td><br />
</tr><br />
</table><br />
<br />
<br />
<br />
<br />
</div><br />
<div><br />
<h2>Discussions on the constants</h2><br />
All the constants given above is steady and theoretically measurable when all the conditions are constant. For example, we could measure k<sub>0</sub> by constructing a new engineered bacteria, which contains the gene encoding SdpC and marker gene alone and observing the influence of the concentration of SdpC on its expression. Yet any modification on genome is notoriously time-consuming, which inhibited us from measuring them in person. We also looked up oceans of papers to confer their approximate ranges, but almost all papers are too fragmental to afford any valid information. Therefore, we decided to assume all these constant according to our limited information and make a qualitative analysis instead of quantifiable analysis. All units and dimensions were temporarily ignored. In other words, our model aims at justifying the validity of this suicide mechanism rather than predicting the exact time or any other parameters of the system.<br />
<br />
Despite the fact that we have hardly any accurate data on these constants, there are some limitations that we extrapolated from known information before we further explore this model:</br><br />
<ol><br />
<li>k<sub>0</sub>>>k<sub>4</sub>≈k<sub>7</sub>: k<sub>0</sub>,k<sub>4</sub> and k<sub>7</sub> represent the normal expression rate of SdpC, SdpR and SdpC separately, and the copy number of SdpC is much larger than that of SdpR and SdpI, whereas the value of the latter two is approximately equal;</li><br />
<li>k<sub>2</sub>>>k<sub>9</sub>: the existence of free SdpR represses the expression of both SdpI and SdpC, and similarly, since the copy number of SdpC is much higher, we expected the repression effect was stronger accordingly;</li><br />
<li>k<sub>10</sub>>>k<sub>3</sub>,k<sub>6</sub>:it is hard to predict the value of k<sub>3</sub> and k<sub>8</sub>, yet we suppose both of them is much smaller than k<sub>10</sub> because SdpI is a kind of membrane protein inherently, and rarely exists as free protein</li><br />
<li>The primary values of all the six variables are very small or strictly zero. We expect it as the most logical initial status. If the primary value of any variable is relatively large, the suicide mechanism may not run normally</li><br />
</ol><br />
</br></br><br />
<h2>Stimulation and discussion</h2><br />
<br />
Simple and rough as the above model is, it does theoretically sound. To test the validity of this model, we first tried to get analytic solution of the ODE set. If this analytic solution exists, we could further investigate the interaction among those variables, and draw some phase planes to get accurate and mathematically perfect description of this model. Unfortunately but expectedly, the existence of analytic solution was negated by MATLAB, and we had to assume groups of values for these constants in advance and analyze the arithmetic solutions instead. These arithmetic solutions not only justified this mechanism is effective enough to commit cell suicide but also indicated some unexpected, or even weird results that beyond our wildest imagination. There are two possibility account for the unexpected results: our model is too rough to include some assignable factor; or there are some implicit but objective limitation inside model, which may be substantiate by later experiments or papers.</br><br />
When we explored the arithmetic solutions of this ODE set, we received nearly one hundred warnings from MATLAB and for many times our most powerful computer ran out of its 8GB memory, but sometimes we can receive the solution within seconds. We had adjusted our parameters for several times before we got our first solution. Here is the values of parameters for this group, and the graph of arithmetic solutions is also given:</br><br />
<div style="margin-top:20px;"><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>50</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table><br />
</div></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/2/26/Suicide1.png"></br></br><br />
At the first glance this graph seemed fine. Initially the concentration of SdpC decreased slightly due to the capturing of SdpI and the repression of float SdpR, but gradually the positive feedback loop works, and C<sub>f</sub> increases rapidly. But when we turned our attention to the curves of other parameters, things seemed not so perfect:</br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/c/cf/Suicide2.png"></br></br><br />
The curve of I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub> contradicted our common sense severely. First, I<sub>m</sub>>C<sub>i</sub>>R<sub>i</sub> is expected to be tenable all the time, which precludes the intersects among the three curves; Second, there is no mechanism in this system that could decrease their concentration, and all of them are increasing function; Third and most serious, never will them be negative, as they represent the concentration of real substances.<br />
<br />
Then we adjusted the parameters slightly for several times. To eliminate those absurd curves, we reconsidered some assumptions.<br />
Here we listed another representative group of parament values and relative graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/7/7d/Suicide3.png/800px-Suicide3.png"></br></br><br />
In this group, we gave up one former assumption and set k<sub>2</sub> equal to k<sub>9</sub>. We also gave positive values to I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub>, which were considered to be zero at first. And by groups of stimulations we realized the value of k<sub>2</sub> does matter, as the derivative of C<sub>f</sub> only increased slightly as k<sub>2</sub> lowers, and the positive values failed to avoid the weird phenomenon in the latter three curves.<br />
We also found that however we adjusted the primary value of I<sub>f</sub> and other parameters, If dropped into approximately zero extremely rapidly at the initial stage and remained balanced, which might account for why the derivatives of the latter curves were abnormally negative. Thus we modified another assumption and increased k<sub>7</sub>. Here is another group of values and corresponding graph:</br></br><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/1/15/Suicide4.png/800px-Suicide4.png" style="width:600; height:400;"></br></br><br />
Although the derivative of Im is not seriously positive constantly, the three latter curves seemed much more reasonable. Hence, we extrapolated although SdpI and SdpR share the same promoter, the expression of SdpI must much faster than SdpR to ensure successful “suicide.” Additionally, the increase of k<sub>7</sub> also represses SdpC, and hence the copy number of SdpC must be larger.<br />
We kept all other parameters constant and gradually augmented k<sub>0</sub>. The larger k<sub>0</sub>, the more perfect the curve seemed, and here are the values table and graph where k<sub>0</sub> equals 400, 80 times larger than k<sub>4</sub>.</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>400</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/3/33/Suicide5.png/800px-Suicide5.png"></br></br><br />
Take the curve of C<sub>f</sub> and R<sub>f</sub> separately, the curves seemed more perfect:</br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/9/9f/Suicide6.png"></br></br><br />
<br />
In wild bacteria who are unable to produced SdpC, naturally k<sub>0</sub> equals zero. We expected C<sub>f</sub> would decreased gradually and finally approximate zero, and here are the corresponding table and graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>30</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/1/19/Suicide7.png"></br></br><br />
Wired but not surprising, there were intersects among the latter three curve, and C<sub>f</sub> decreases continually when it is negative. We continued to try groups of these parameters, and this is the best one where we increased the primary concentration of SdpC and the normal expression rate of SdpI.<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/8/8e/Suicide8.png"></br></br><br />
The curve of C<sub>f</sub> and R<sub>f</sub> alone:<br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/e/eb/Suicide9.png"></br></br><br />
In spite of minimal abnormal phenomenon (C<sub>f</sub> was negative in later stage), this graph roughly testified that in wild bacterial the concentration of float SdpC will drop to nearly zero quickly.<br />
In sum, the ODE model of singular cells indicate following results:</br><br />
<ol><br />
<li>The character of free SdpC is most affected by k<sub>0</sub>, if the copy number of SdpC is large enough, it is theoretically reasonable to commit suicide;</li><br />
<li>The influence of the value of Imax and k<sub>2</sub> is much limited;</li><br />
<li>The amount of free SdpI is always near zero;</li><br />
<li>SdpC will not increase limitlessly however we transform parameters;</li><br />
<li>To ensure the success of suicide, it is required k<sub>0</sub>>>k<sub>4</sub>>>k<sub>7</sub>;</li><br />
The last conclusion was our biggest windfall, and we have verified the validity of this suicide mechanism in math. On the one hand, if further experiments proven #4 engineered bacteria will kill both siblings and themselves, it is highly like that the expression rate SdpI is much larger than SdpR even if they share the same promoter; on the other hand, if #4 engineered bacteria are not able to commit suicide, we can try to boost the expression of SdpI to adjust the bacteria.<br />
<br />
<br />
<h1>Discussion on colonies</h1><br />
<br />
In reality, the engineered bacteria aims at killing its siblings instead of itself, and at first almost all toxin SdpC will be secreted outside the bacteria. Assume the diffusion of toxin among cells comply with diffusion law, that is, the diffusion rate is proportionate with the gradient of concentration. Further assume the death concentration of SdpC is same to all bacteria expect those who contain this locus, the average life expectancy is bacteria will hinge on the rate and distribution of engineered bacteria, and the distribution of life expectancy of bacteria is similar to that of average free path of gas molecules.<br />
As long the coefficient of diffusion is large enough, any engineered bacterias, no matter how few, is adequate to devastate the whole colony. Alike to the average free path of thin gas, the average suicide time of the whole reporter system is inversely proportional with the square root of the rate of engineer bacteria containing this locus.<br />
<br />
<br />
<h1>References</h1><br />
Parallel pathways of repression and antirepression governing the transition to stationary phase in Bacillus subtilis<br />
AV Banse, A Chastanet, L Rahn-Lee…,PNAS ,2008 </br><br />
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<h1>Introduction</h1><br />
To be more user-friendly, 4# circuit contains a reporting system. After melting in water, the spores germinate and express blue pigment protein to report the best using time.</br><br />
Meanwhile, 4# circuit could also ensure biosafety. Because other circuit do not have self-killing device, 4# engineering bacterial should kill all the bacterial after using.</br><br />
<br />
</div><br />
<br />
<div><br />
<h1>Designing of the suicide system</h1><br />
We design a circuit of killing switch based on its endogenous genetic system.</br><br />
In B.subtilis, when it comes to the stationary phase, the environmental pressure increases and nutrition becomes limited, so B begin to produce spores. Now the community will be divided into two different parts. One of them are trying to kill others to get enough nutrient , delaying the production of spores and achieving a competitive advantage. Killing is mediated by the exported toxic protein SdpC. SdpI will appear on the membrane surface to avoid itself from being damaged. SdpI could bind free SdpC and autopressor SdpR, to remove SdpR’s inhibition against I and R, to produce more SdpI to offset SdpC, finally guaranteeing the subgroup alive, thereby delaying the spores production.</br><br />
<img src="https://static.igem.org/mediawiki/2013/2/2b/Reporter_3.png" width="500" height="350"><br />
<p>We transfer SdpC which is fused by promoter SdpI/R into high copy plasmids in order to damage the balance of the system, thereby killing whole colony. When SdpC appears, SdpI on the membrane will bind free SdpC and adsorb SdpR to cease its inhibition against SdpI P/R, trying to produce more SdpI. At the same time, it will activate the promoter SdpR/I in our circuits and generate more SdpC.The system would fall into an infinite loop, and according to our modeling ,the amount of SdpC increases beyond the ability of SdpI.Thus,the cells with protection mechanism will crack and die because of too much SdpC. All above forms the killing device.<br />
We Also designed a test circuit,which contains promotor grac and sdpABC only,aiming to determine the ability of SdpC.</p><br />
<br />
<div></br></br><br />
There are both positive and negative feedback loops in this process. On the one hand, SdpI is unable to sequestrate the autorepressor, SdpR, until it captures the toxin, SdpC. The accumulation of SdpC will thus facilitate SdpI to capture more SdpR and thereby relieve the repression of SdpR, stimulating the expression of itself. This is the positive feedback loop which leads to the increasing accumulation of SdpC and finally the death of the bacteria. On the other hand, the removal of SdpR also enhance the expression of SdpI and accelerate the sequestration of SdpC, which forms a negative feedback loop whose effects contradict the positive feedback loop. However, since the copy number of SdpC is much higher, it is believed that the positive loop is strong enough to outweigh the negative one, which guarantees this mechanism will finally leads to collapse instead of equilibrium.<br />
</div><br />
<div><br />
<h1>The ODE model of singular cells</h1><br />
There is no denying fact that the essential goal of engineered bacterias who carry this so called “suicide” locus itself is to kill their siblings rather than themselves to ensure the survival of themselves. Surly they can kill their siblings, but can they finally eliminate themselves, as we expects? The trivial experiment protocol and huge uncertainty had put off our experiment, and as expected, we failed to achieve the construction of complete reporter system in our laboratory. Fortunately, we could resort to mathematical models to verify the validity of this locus theoretically. <br />
There are six independent variables in individual cells, and the theoretically if the initial conditions are fixed, all of them will be the univariate functions of time. The following table illustrates the mark and meaning of each variable:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub></td><br />
<td >Mole number of free SdpI in cytoplasm</a>.</td><br />
</tr><br />
<tr><br />
<td >I<sub>m</sub> </td><br />
<td >Mole number of SdpI in the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >C<sub>f</sub></td><br />
<td >Mole number of free SdpC in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >C<sub>i</sub></td><br />
<td >Mole number of SdpC captured by SdpI.</td><br />
</tr><br />
<tr><br />
<td >R<sub>f</sub></td><br />
<td >Mole number of free SdpR in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >R<sub>i</sub> </td><br />
<td >Mole number of SdpR captured by SdpI</td><br />
</tr><br />
</table><br />
</br></br><br />
<div>To construct reasonable ordinary differential equation (ODE) model to describe and predict the operation of the suicide system, we followed the law of mass action, one basic law of chemistry and biology.</br><br />
Taken as a statement about kinetics, the law states that the rate of an elementary reaction (a reaction that proceeds through only one transition state, which is one mechanistic step) is proportional to the product of the concentrations of the participating molecules. In modern chemistry this is derived using statistical mechanics. Despite the complicated chemical reactions involved in the process of transcription and translation, it is common and logically sound to view the expression of one particular gene as an elementary reaction and assume the repression effects of the protein itself encodes and the repressor are both linear.</br><br />
According to the law of mass action, we got six independent differential equation of the variables:</br><br />
<img src="https://static.igem.org/mediawiki/2013/e/e8/For1%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/2/23/For2%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/4/48/For3%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/7/71/For4%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/3/31/For5%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/c/cb/For6%281%29.png"></br></br></br><br />
The following table explain the constants in the above ODE groups:</br></br></div><br />
<div style="margin-top:20px;"><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub> </td><br />
<td >The maximal number of SdpI than can be fixed on the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >k<sub>0</sub></td><br />
<td >Constant describes the normal expression rate of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>1</sub> </td><br />
<td >Constant describes the self-repression effects of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>2</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpC.</a></td><br />
</tr><br />
<tr><br />
<td >k<sub>3</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>4</sub> </td><br />
<td >Constant describes the normal expression rate of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>5</sub> </td><br />
<td >Constant describes the self-repression effects of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>6</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>7</sub> </td><br />
<td >Constant describes the normal expression rate of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>8</sub> </td><br />
<td >Constant describes the self-repression effects of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>9</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>10</sub> </td><br />
<td >Constant describes the rate of SdpI binding to the cell membrane</td><br />
</tr><br />
</table><br />
</div><br />
<br />
<br />
<br />
<br />
</div><br />
<div><br />
<h2>Discussions on the constants</h2><br />
All the constants given above is steady and theoretically measurable when all the conditions are constant. For example, we could measure k<sub>0</sub> by constructing a new engineered bacteria, which contains the gene encoding SdpC and marker gene alone and observing the influence of the concentration of SdpC on its expression. Yet any modification on genome is notoriously time-consuming, which inhibited us from measuring them in person. We also looked up oceans of papers to confer their approximate ranges, but almost all papers are too fragmental to afford any valid information. Therefore, we decided to assume all these constant according to our limited information and make a qualitative analysis instead of quantifiable analysis. All units and dimensions were temporarily ignored. In other words, our model aims at justifying the validity of this suicide mechanism rather than predicting the exact time or any other parameters of the system.<br />
<br />
Despite the fact that we have hardly any accurate data on these constants, there are some limitations that we extrapolated from known information before we further explore this model:</br><br />
<ol><br />
<li>k<sub>0</sub>>>k<sub>4</sub>≈k<sub>7</sub>: k<sub>0</sub>,k<sub>4</sub> and k<sub>7</sub> represent the normal expression rate of SdpC, SdpR and SdpC separately, and the copy number of SdpC is much larger than that of SdpR and SdpI, whereas the value of the latter two is approximately equal;</li><br />
<li>k<sub>2</sub>>>k<sub>9</sub>: the existence of free SdpR represses the expression of both SdpI and SdpC, and similarly, since the copy number of SdpC is much higher, we expected the repression effect was stronger accordingly;</li><br />
<li>k<sub>10</sub>>>k<sub>3</sub>,k<sub>6</sub>:it is hard to predict the value of k<sub>3</sub> and k<sub>8</sub>, yet we suppose both of them is much smaller than k<sub>10</sub> because SdpI is a kind of membrane protein inherently, and rarely exists as free protein</li><br />
<li>The primary values of all the six variables are very small or strictly zero. We expect it as the most logical initial status. If the primary value of any variable is relatively large, the suicide mechanism may not run normally</li><br />
</ol><br />
</br></br><br />
<h2>Stimulation and discussion</h2><br />
<br />
Simple and rough as the above model is, it does theoretically sound. To test the validity of this model, we first tried to get analytic solution of the ODE set. If this analytic solution exists, we could further investigate the interaction among those variables, and draw some phase planes to get accurate and mathematically perfect description of this model. Unfortunately but expectedly, the existence of analytic solution was negated by MATLAB, and we had to assume groups of values for these constants in advance and analyze the arithmetic solutions instead. These arithmetic solutions not only justified this mechanism is effective enough to commit cell suicide but also indicated some unexpected, or even weird results that beyond our wildest imagination. There are two possibility account for the unexpected results: our model is too rough to include some assignable factor; or there are some implicit but objective limitation inside model, which may be substantiate by later experiments or papers.</br><br />
When we explored the arithmetic solutions of this ODE set, we received nearly one hundred warnings from MATLAB and for many times our most powerful computer ran out of its 8GB memory, but sometimes we can receive the solution within seconds. We had adjusted our parameters for several times before we got our first solution. Here is the values of parameters for this group, and the graph of arithmetic solutions is also given:</br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>50</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/2/26/Suicide1.png"></br></br><br />
At the first glance this graph seemed fine. Initially the concentration of SdpC decreased slightly due to the capturing of SdpI and the repression of float SdpR, but gradually the positive feedback loop works, and C<sub>f</sub> increases rapidly. But when we turned our attention to the curves of other parameters, things seemed not so perfect:</br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/c/cf/Suicide2.png"></br></br><br />
The curve of I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub> contradicted our common sense severely. First, I<sub>m</sub>>C<sub>i</sub>>R<sub>i</sub> is expected to be tenable all the time, which precludes the intersects among the three curves; Second, there is no mechanism in this system that could decrease their concentration, and all of them are increasing function; Third and most serious, never will them be negative, as they represent the concentration of real substances.<br />
<br />
Then we adjusted the parameters slightly for several times. To eliminate those absurd curves, we reconsidered some assumptions.<br />
Here we listed another representative group of parament values and relative graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/7/7d/Suicide3.png/800px-Suicide3.png"></br></br><br />
In this group, we gave up one former assumption and set k<sub>2</sub> equal to k<sub>9</sub>. We also gave positive values to I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub>, which were considered to be zero at first. And by groups of stimulations we realized the value of k<sub>2</sub> does matter, as the derivative of C<sub>f</sub> only increased slightly as k<sub>2</sub> lowers, and the positive values failed to avoid the weird phenomenon in the latter three curves.<br />
We also found that however we adjusted the primary value of I<sub>f</sub> and other parameters, If dropped into approximately zero extremely rapidly at the initial stage and remained balanced, which might account for why the derivatives of the latter curves were abnormally negative. Thus we modified another assumption and increased k<sub>7</sub>. Here is another group of values and corresponding graph:</br></br><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/1/15/Suicide4.png/800px-Suicide4.png" style="width:600; height:400;"></br></br><br />
Although the derivative of Im is not seriously positive constantly, the three latter curves seemed much more reasonable. Hence, we extrapolated although SdpI and SdpR share the same promoter, the expression of SdpI must much faster than SdpR to ensure successful “suicide.” Additionally, the increase of k<sub>7</sub> also represses SdpC, and hence the copy number of SdpC must be larger.<br />
We kept all other parameters constant and gradually augmented k<sub>0</sub>. The larger k<sub>0</sub>, the more perfect the curve seemed, and here are the values table and graph where k<sub>0</sub> equals 400, 80 times larger than k<sub>4</sub>.</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>400</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/3/33/Suicide5.png/800px-Suicide5.png"></br></br><br />
Take the curve of C<sub>f</sub> and R<sub>f</sub> separately, the curves seemed more perfect:</br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/9/9f/Suicide6.png"></br></br><br />
<br />
In wild bacteria who are unable to produced SdpC, naturally k<sub>0</sub> equals zero. We expected C<sub>f</sub> would decreased gradually and finally approximate zero, and here are the corresponding table and graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>30</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/1/19/Suicide7.png"></br></br><br />
Wired but not surprising, there were intersects among the latter three curve, and C<sub>f</sub> decreases continually when it is negative. We continued to try groups of these parameters, and this is the best one where we increased the primary concentration of SdpC and the normal expression rate of SdpI.<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/8/8e/Suicide8.png"></br></br><br />
The curve of C<sub>f</sub> and R<sub>f</sub> alone:<br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/e/eb/Suicide9.png"></br></br><br />
In spite of minimal abnormal phenomenon (C<sub>f</sub> was negative in later stage), this graph roughly testified that in wild bacterial the concentration of float SdpC will drop to nearly zero quickly.<br />
In sum, the ODE model of singular cells indicate following results:</br><br />
<ol><br />
<li>The character of free SdpC is most affected by k<sub>0</sub>, if the copy number of SdpC is large enough, it is theoretically reasonable to commit suicide;</li><br />
<li>The influence of the value of Imax and k<sub>2</sub> is much limited;</li><br />
<li>The amount of free SdpI is always near zero;</li><br />
<li>SdpC will not increase limitlessly however we transform parameters;</li><br />
<li>To ensure the success of suicide, it is required k<sub>0</sub>>>k<sub>4</sub>>>k<sub>7</sub>;</li><br />
The last conclusion was our biggest windfall, and we have verified the validity of this suicide mechanism in math. On the one hand, if further experiments proven #4 engineered bacteria will kill both siblings and themselves, it is highly like that the expression rate SdpI is much larger than SdpR even if they share the same promoter; on the other hand, if #4 engineered bacteria are not able to commit suicide, we can try to boost the expression of SdpI to adjust the bacteria.<br />
<br />
<br />
<h1>Discussion on colonies</h1><br />
<br />
In reality, the engineered bacteria aims at killing its siblings instead of itself, and at first almost all toxin SdpC will be secreted outside the bacteria. Assume the diffusion of toxin among cells comply with diffusion law, that is, the diffusion rate is proportionate with the gradient of concentration. Further assume the death concentration of SdpC is same to all bacteria expect those who contain this locus, the average life expectancy is bacteria will hinge on the rate and distribution of engineered bacteria, and the distribution of life expectancy of bacteria is similar to that of average free path of gas molecules.<br />
As long the coefficient of diffusion is large enough, any engineered bacterias, no matter how few, is adequate to devastate the whole colony. Alike to the average free path of thin gas, the average suicide time of the whole reporter system is inversely proportional with the square root of the rate of engineer bacteria containing this locus.<br />
<br />
<br />
<h1>References</h1><br />
Parallel pathways of repression and antirepression governing the transition to stationary phase in Bacillus subtilis<br />
AV Banse, A Chastanet, L Rahn-Lee…,PNAS ,2008 </br><br />
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<h1>Introduction</h1><br />
To be more user-friendly, 4# circuit contains a reporting system. After melting in water, the spores germinate and express blue pigment protein to report the best using time.</br><br />
Meanwhile, 4# circuit could also ensure biosafety. Because other circuit do not have self-killing device, 4# engineering bacterial should kill all the bacterial after using.</br><br />
<br />
</div><br />
<br />
<div><br />
<h1>Designing of the suicide system</h1><br />
We design a circuit of killing switch based on its endogenous genetic system.</br><br />
In B.subtilis, when it comes to the stationary phase, the environmental pressure increases and nutrition becomes limited, so B begin to produce spores. Now the community will be divided into two different parts. One of them are trying to kill others to get enough nutrient , delaying the production of spores and achieving a competitive advantage. Killing is mediated by the exported toxic protein SdpC. SdpI will appear on the membrane surface to avoid itself from being damaged. SdpI could bind free SdpC and autopressor SdpR, to remove SdpR’s inhibition against I and R, to produce more SdpI to offset SdpC, finally guaranteeing the subgroup alive, thereby delaying the spores production.</br><br />
<img src="https://static.igem.org/mediawiki/2013/2/2b/Reporter_3.png" width="500" height="350"><br />
<p>We transfer SdpC which is fused by promoter SdpI/R into high copy plasmids in order to damage the balance of the system, thereby killing whole colony. When SdpC appears, SdpI on the membrane will bind free SdpC and adsorb SdpR to cease its inhibition against SdpI P/R, trying to produce more SdpI. At the same time, it will activate the promoter SdpR/I in our circuits and generate more SdpC.The system would fall into an infinite loop, and according to our modeling ,the amount of SdpC increases beyond the ability of SdpI.Thus,the cells with protection mechanism will crack and die because of too much SdpC. All above forms the killing device.<br />
We Also designed a test circuit,which contains promotor grac and sdpABC only,aiming to determine the ability of SdpC.</p><br />
<br />
<div></br></br><br />
There are both positive and negative feedback loops in this process. On the one hand, SdpI is unable to sequestrate the autorepressor, SdpR, until it captures the toxin, SdpC. The accumulation of SdpC will thus facilitate SdpI to capture more SdpR and thereby relieve the repression of SdpR, stimulating the expression of itself. This is the positive feedback loop which leads to the increasing accumulation of SdpC and finally the death of the bacteria. On the other hand, the removal of SdpR also enhance the expression of SdpI and accelerate the sequestration of SdpC, which forms a negative feedback loop whose effects contradict the positive feedback loop. However, since the copy number of SdpC is much higher, it is believed that the positive loop is strong enough to outweigh the negative one, which guarantees this mechanism will finally leads to collapse instead of equilibrium.<br />
</div><br />
<div><br />
<h1>The ODE model of singular cells</h1><br />
There is no denying fact that the essential goal of engineered bacterias who carry this so called “suicide” locus itself is to kill their siblings rather than themselves to ensure the survival of themselves. Surly they can kill their siblings, but can they finally eliminate themselves, as we expects? The trivial experiment protocol and huge uncertainty had put off our experiment, and as expected, we failed to achieve the construction of complete reporter system in our laboratory. Fortunately, we could resort to mathematical models to verify the validity of this locus theoretically. <br />
There are six independent variables in individual cells, and the theoretically if the initial conditions are fixed, all of them will be the univariate functions of time. The following table illustrates the mark and meaning of each variable:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub></td><br />
<td >Mole number of free SdpI in cytoplasm</a>.</td><br />
</tr><br />
<tr><br />
<td >I<sub>m</sub> </td><br />
<td >Mole number of SdpI in the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >C<sub>f</sub></td><br />
<td >Mole number of free SdpC in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >C<sub>i</sub></td><br />
<td >Mole number of SdpC captured by SdpI.</td><br />
</tr><br />
<tr><br />
<td >R<sub>f</sub></td><br />
<td >Mole number of free SdpR in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >R<sub>i</sub> </td><br />
<td >Mole number of SdpR captured by SdpI</td><br />
</tr><br />
</table><br />
</br></br><br />
To construct reasonable ordinary differential equation (ODE) model to describe and predict the operation of the suicide system, we followed the law of mass action, one basic law of chemistry and biology.</br><br />
Taken as a statement about kinetics, the law states that the rate of an elementary reaction (a reaction that proceeds through only one transition state, which is one mechanistic step) is proportional to the product of the concentrations of the participating molecules. In modern chemistry this is derived using statistical mechanics. Despite the complicated chemical reactions involved in the process of transcription and translation, it is common and logically sound to view the expression of one particular gene as an elementary reaction and assume the repression effects of the protein itself encodes and the repressor are both linear.</br><br />
According to the law of mass action, we got six independent differential equation of the variables:</br><br />
<img src="https://static.igem.org/mediawiki/2013/e/e8/For1%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/2/23/For2%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/4/48/For3%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/7/71/For4%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/3/31/For5%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/c/cb/For6%281%29.png"></br></br></br><br />
The following table explain the constants in the above ODE groups:</br></br><br />
<div style="width:580px;height:20px;"></div><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub> </td><br />
<td >The maximal number of SdpI than can be fixed on the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >k<sub>0</sub></td><br />
<td >Constant describes the normal expression rate of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>1</sub> </td><br />
<td >Constant describes the self-repression effects of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>2</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpC.</a></td><br />
</tr><br />
<tr><br />
<td >k<sub>3</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>4</sub> </td><br />
<td >Constant describes the normal expression rate of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>5</sub> </td><br />
<td >Constant describes the self-repression effects of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>6</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>7</sub> </td><br />
<td >Constant describes the normal expression rate of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>8</sub> </td><br />
<td >Constant describes the self-repression effects of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>9</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>10</sub> </td><br />
<td >Constant describes the rate of SdpI binding to the cell membrane</td><br />
</tr><br />
</table><br />
<br />
<br />
<br />
<br />
</div><br />
<div><br />
<h2>Discussions on the constants</h2><br />
All the constants given above is steady and theoretically measurable when all the conditions are constant. For example, we could measure k<sub>0</sub> by constructing a new engineered bacteria, which contains the gene encoding SdpC and marker gene alone and observing the influence of the concentration of SdpC on its expression. Yet any modification on genome is notoriously time-consuming, which inhibited us from measuring them in person. We also looked up oceans of papers to confer their approximate ranges, but almost all papers are too fragmental to afford any valid information. Therefore, we decided to assume all these constant according to our limited information and make a qualitative analysis instead of quantifiable analysis. All units and dimensions were temporarily ignored. In other words, our model aims at justifying the validity of this suicide mechanism rather than predicting the exact time or any other parameters of the system.<br />
<br />
Despite the fact that we have hardly any accurate data on these constants, there are some limitations that we extrapolated from known information before we further explore this model:</br><br />
<ol><br />
<li>k<sub>0</sub>>>k<sub>4</sub>≈k<sub>7</sub>: k<sub>0</sub>,k<sub>4</sub> and k<sub>7</sub> represent the normal expression rate of SdpC, SdpR and SdpC separately, and the copy number of SdpC is much larger than that of SdpR and SdpI, whereas the value of the latter two is approximately equal;</li><br />
<li>k<sub>2</sub>>>k<sub>9</sub>: the existence of free SdpR represses the expression of both SdpI and SdpC, and similarly, since the copy number of SdpC is much higher, we expected the repression effect was stronger accordingly;</li><br />
<li>k<sub>10</sub>>>k<sub>3</sub>,k<sub>6</sub>:it is hard to predict the value of k<sub>3</sub> and k<sub>8</sub>, yet we suppose both of them is much smaller than k<sub>10</sub> because SdpI is a kind of membrane protein inherently, and rarely exists as free protein</li><br />
<li>The primary values of all the six variables are very small or strictly zero. We expect it as the most logical initial status. If the primary value of any variable is relatively large, the suicide mechanism may not run normally</li><br />
</ol><br />
</br></br><br />
<h2>Stimulation and discussion</h2><br />
<br />
Simple and rough as the above model is, it does theoretically sound. To test the validity of this model, we first tried to get analytic solution of the ODE set. If this analytic solution exists, we could further investigate the interaction among those variables, and draw some phase planes to get accurate and mathematically perfect description of this model. Unfortunately but expectedly, the existence of analytic solution was negated by MATLAB, and we had to assume groups of values for these constants in advance and analyze the arithmetic solutions instead. These arithmetic solutions not only justified this mechanism is effective enough to commit cell suicide but also indicated some unexpected, or even weird results that beyond our wildest imagination. There are two possibility account for the unexpected results: our model is too rough to include some assignable factor; or there are some implicit but objective limitation inside model, which may be substantiate by later experiments or papers.</br><br />
When we explored the arithmetic solutions of this ODE set, we received nearly one hundred warnings from MATLAB and for many times our most powerful computer ran out of its 8GB memory, but sometimes we can receive the solution within seconds. We had adjusted our parameters for several times before we got our first solution. Here is the values of parameters for this group, and the graph of arithmetic solutions is also given:</br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>50</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/2/26/Suicide1.png"></br></br><br />
At the first glance this graph seemed fine. Initially the concentration of SdpC decreased slightly due to the capturing of SdpI and the repression of float SdpR, but gradually the positive feedback loop works, and C<sub>f</sub> increases rapidly. But when we turned our attention to the curves of other parameters, things seemed not so perfect:</br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/c/cf/Suicide2.png"></br></br><br />
The curve of I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub> contradicted our common sense severely. First, I<sub>m</sub>>C<sub>i</sub>>R<sub>i</sub> is expected to be tenable all the time, which precludes the intersects among the three curves; Second, there is no mechanism in this system that could decrease their concentration, and all of them are increasing function; Third and most serious, never will them be negative, as they represent the concentration of real substances.<br />
<br />
Then we adjusted the parameters slightly for several times. To eliminate those absurd curves, we reconsidered some assumptions.<br />
Here we listed another representative group of parament values and relative graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/7/7d/Suicide3.png/800px-Suicide3.png"></br></br><br />
In this group, we gave up one former assumption and set k<sub>2</sub> equal to k<sub>9</sub>. We also gave positive values to I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub>, which were considered to be zero at first. And by groups of stimulations we realized the value of k<sub>2</sub> does matter, as the derivative of C<sub>f</sub> only increased slightly as k<sub>2</sub> lowers, and the positive values failed to avoid the weird phenomenon in the latter three curves.<br />
We also found that however we adjusted the primary value of I<sub>f</sub> and other parameters, If dropped into approximately zero extremely rapidly at the initial stage and remained balanced, which might account for why the derivatives of the latter curves were abnormally negative. Thus we modified another assumption and increased k<sub>7</sub>. Here is another group of values and corresponding graph:</br></br><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/1/15/Suicide4.png/800px-Suicide4.png" style="width:600; height:400;"></br></br><br />
Although the derivative of Im is not seriously positive constantly, the three latter curves seemed much more reasonable. Hence, we extrapolated although SdpI and SdpR share the same promoter, the expression of SdpI must much faster than SdpR to ensure successful “suicide.” Additionally, the increase of k<sub>7</sub> also represses SdpC, and hence the copy number of SdpC must be larger.<br />
We kept all other parameters constant and gradually augmented k<sub>0</sub>. The larger k<sub>0</sub>, the more perfect the curve seemed, and here are the values table and graph where k<sub>0</sub> equals 400, 80 times larger than k<sub>4</sub>.</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>400</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/3/33/Suicide5.png/800px-Suicide5.png"></br></br><br />
Take the curve of C<sub>f</sub> and R<sub>f</sub> separately, the curves seemed more perfect:</br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/9/9f/Suicide6.png"></br></br><br />
<br />
In wild bacteria who are unable to produced SdpC, naturally k<sub>0</sub> equals zero. We expected C<sub>f</sub> would decreased gradually and finally approximate zero, and here are the corresponding table and graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>30</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/1/19/Suicide7.png"></br></br><br />
Wired but not surprising, there were intersects among the latter three curve, and C<sub>f</sub> decreases continually when it is negative. We continued to try groups of these parameters, and this is the best one where we increased the primary concentration of SdpC and the normal expression rate of SdpI.<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/8/8e/Suicide8.png"></br></br><br />
The curve of C<sub>f</sub> and R<sub>f</sub> alone:<br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/e/eb/Suicide9.png"></br></br><br />
In spite of minimal abnormal phenomenon (C<sub>f</sub> was negative in later stage), this graph roughly testified that in wild bacterial the concentration of float SdpC will drop to nearly zero quickly.<br />
In sum, the ODE model of singular cells indicate following results:</br><br />
<ol><br />
<li>The character of free SdpC is most affected by k<sub>0</sub>, if the copy number of SdpC is large enough, it is theoretically reasonable to commit suicide;</li><br />
<li>The influence of the value of Imax and k<sub>2</sub> is much limited;</li><br />
<li>The amount of free SdpI is always near zero;</li><br />
<li>SdpC will not increase limitlessly however we transform parameters;</li><br />
<li>To ensure the success of suicide, it is required k<sub>0</sub>>>k<sub>4</sub>>>k<sub>7</sub>;</li><br />
The last conclusion was our biggest windfall, and we have verified the validity of this suicide mechanism in math. On the one hand, if further experiments proven #4 engineered bacteria will kill both siblings and themselves, it is highly like that the expression rate SdpI is much larger than SdpR even if they share the same promoter; on the other hand, if #4 engineered bacteria are not able to commit suicide, we can try to boost the expression of SdpI to adjust the bacteria.<br />
<br />
<br />
<h1>Discussion on colonies</h1><br />
<br />
In reality, the engineered bacteria aims at killing its siblings instead of itself, and at first almost all toxin SdpC will be secreted outside the bacteria. Assume the diffusion of toxin among cells comply with diffusion law, that is, the diffusion rate is proportionate with the gradient of concentration. Further assume the death concentration of SdpC is same to all bacteria expect those who contain this locus, the average life expectancy is bacteria will hinge on the rate and distribution of engineered bacteria, and the distribution of life expectancy of bacteria is similar to that of average free path of gas molecules.<br />
As long the coefficient of diffusion is large enough, any engineered bacterias, no matter how few, is adequate to devastate the whole colony. Alike to the average free path of thin gas, the average suicide time of the whole reporter system is inversely proportional with the square root of the rate of engineer bacteria containing this locus.<br />
<br />
<br />
<h1>References</h1><br />
Parallel pathways of repression and antirepression governing the transition to stationary phase in Bacillus subtilis<br />
AV Banse, A Chastanet, L Rahn-Lee…,PNAS ,2008 </br><br />
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<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a>&gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></div></div><br />
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<h1>Introduction</h1><br />
To be more user-friendly, 4# circuit contains a reporting system. After melting in water, the spores germinate and express blue pigment protein to report the best using time.</br><br />
Meanwhile, 4# circuit could also ensure biosafety. Because other circuit do not have self-killing device, 4# engineering bacterial should kill all the bacterial after using.</br><br />
<br />
</div><br />
<br />
<div><br />
<h1>Designing of the suicide system</h1><br />
We design a circuit of killing switch based on its endogenous genetic system.</br><br />
In B.subtilis, when it comes to the stationary phase, the environmental pressure increases and nutrition becomes limited, so B begin to produce spores. Now the community will be divided into two different parts. One of them are trying to kill others to get enough nutrient , delaying the production of spores and achieving a competitive advantage. Killing is mediated by the exported toxic protein SdpC. SdpI will appear on the membrane surface to avoid itself from being damaged. SdpI could bind free SdpC and autopressor SdpR, to remove SdpR’s inhibition against I and R, to produce more SdpI to offset SdpC, finally guaranteeing the subgroup alive, thereby delaying the spores production.</br><br />
<img src="https://static.igem.org/mediawiki/2013/2/2b/Reporter_3.png" width="500" height="350"><br />
<p>We transfer SdpC which is fused by promoter SdpI/R into high copy plasmids in order to damage the balance of the system, thereby killing whole colony. When SdpC appears, SdpI on the membrane will bind free SdpC and adsorb SdpR to cease its inhibition against SdpI P/R, trying to produce more SdpI. At the same time, it will activate the promoter SdpR/I in our circuits and generate more SdpC.The system would fall into an infinite loop, and according to our modeling ,the amount of SdpC increases beyond the ability of SdpI.Thus,the cells with protection mechanism will crack and die because of too much SdpC. All above forms the killing device.<br />
We Also designed a test circuit,which contains promotor grac and sdpABC only,aiming to determine the ability of SdpC.</p><br />
<br />
<div></br></br><br />
There are both positive and negative feedback loops in this process. On the one hand, SdpI is unable to sequestrate the autorepressor, SdpR, until it captures the toxin, SdpC. The accumulation of SdpC will thus facilitate SdpI to capture more SdpR and thereby relieve the repression of SdpR, stimulating the expression of itself. This is the positive feedback loop which leads to the increasing accumulation of SdpC and finally the death of the bacteria. On the other hand, the removal of SdpR also enhance the expression of SdpI and accelerate the sequestration of SdpC, which forms a negative feedback loop whose effects contradict the positive feedback loop. However, since the copy number of SdpC is much higher, it is believed that the positive loop is strong enough to outweigh the negative one, which guarantees this mechanism will finally leads to collapse instead of equilibrium.<br />
</div><br />
<div><br />
<h1>The ODE model of singular cells</h1><br />
There is no denying fact that the essential goal of engineered bacterias who carry this so called “suicide” locus itself is to kill their siblings rather than themselves to ensure the survival of themselves. Surly they can kill their siblings, but can they finally eliminate themselves, as we expects? The trivial experiment protocol and huge uncertainty had put off our experiment, and as expected, we failed to achieve the construction of complete reporter system in our laboratory. Fortunately, we could resort to mathematical models to verify the validity of this locus theoretically. <br />
There are six independent variables in individual cells, and the theoretically if the initial conditions are fixed, all of them will be the univariate functions of time. The following table illustrates the mark and meaning of each variable:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub></td><br />
<td >Mole number of free SdpI in cytoplasm</a>.</td><br />
</tr><br />
<tr><br />
<td >I<sub>m</sub> </td><br />
<td >Mole number of SdpI in the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >C<sub>f</sub></td><br />
<td >Mole number of free SdpC in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >C<sub>i</sub></td><br />
<td >Mole number of SdpC captured by SdpI.</td><br />
</tr><br />
<tr><br />
<td >R<sub>f</sub></td><br />
<td >Mole number of free SdpR in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >R<sub>i</sub> </td><br />
<td >Mole number of SdpR captured by SdpI</td><br />
</tr><br />
</table><br />
</br></br><br />
To construct reasonable ordinary differential equation (ODE) model to describe and predict the operation of the suicide system, we followed the law of mass action, one basic law of chemistry and biology.</br><br />
Taken as a statement about kinetics, the law states that the rate of an elementary reaction (a reaction that proceeds through only one transition state, which is one mechanistic step) is proportional to the product of the concentrations of the participating molecules. In modern chemistry this is derived using statistical mechanics. Despite the complicated chemical reactions involved in the process of transcription and translation, it is common and logically sound to view the expression of one particular gene as an elementary reaction and assume the repression effects of the protein itself encodes and the repressor are both linear.</br><br />
According to the law of mass action, we got six independent differential equation of the variables:</br><br />
<img src="https://static.igem.org/mediawiki/2013/e/e8/For1%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/2/23/For2%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/4/48/For3%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/7/71/For4%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/3/31/For5%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/c/cb/For6%281%29.png"></br></br></br><br />
The following table explain the constants in the above ODE groups:</br></br><br />
<p></p><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub> </td><br />
<td >The maximal number of SdpI than can be fixed on the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >k<sub>0</sub></td><br />
<td >Constant describes the normal expression rate of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>1</sub> </td><br />
<td >Constant describes the self-repression effects of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>2</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpC.</a></td><br />
</tr><br />
<tr><br />
<td >k<sub>3</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>4</sub> </td><br />
<td >Constant describes the normal expression rate of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>5</sub> </td><br />
<td >Constant describes the self-repression effects of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>6</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>7</sub> </td><br />
<td >Constant describes the normal expression rate of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>8</sub> </td><br />
<td >Constant describes the self-repression effects of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>9</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>10</sub> </td><br />
<td >Constant describes the rate of SdpI binding to the cell membrane</td><br />
</tr><br />
</table><br />
<br />
<br />
<br />
<br />
</div><br />
<div><br />
<h2>Discussions on the constants</h2><br />
All the constants given above is steady and theoretically measurable when all the conditions are constant. For example, we could measure k<sub>0</sub> by constructing a new engineered bacteria, which contains the gene encoding SdpC and marker gene alone and observing the influence of the concentration of SdpC on its expression. Yet any modification on genome is notoriously time-consuming, which inhibited us from measuring them in person. We also looked up oceans of papers to confer their approximate ranges, but almost all papers are too fragmental to afford any valid information. Therefore, we decided to assume all these constant according to our limited information and make a qualitative analysis instead of quantifiable analysis. All units and dimensions were temporarily ignored. In other words, our model aims at justifying the validity of this suicide mechanism rather than predicting the exact time or any other parameters of the system.<br />
<br />
Despite the fact that we have hardly any accurate data on these constants, there are some limitations that we extrapolated from known information before we further explore this model:</br><br />
<ol><br />
<li>k<sub>0</sub>>>k<sub>4</sub>≈k<sub>7</sub>: k<sub>0</sub>,k<sub>4</sub> and k<sub>7</sub> represent the normal expression rate of SdpC, SdpR and SdpC separately, and the copy number of SdpC is much larger than that of SdpR and SdpI, whereas the value of the latter two is approximately equal;</li><br />
<li>k<sub>2</sub>>>k<sub>9</sub>: the existence of free SdpR represses the expression of both SdpI and SdpC, and similarly, since the copy number of SdpC is much higher, we expected the repression effect was stronger accordingly;</li><br />
<li>k<sub>10</sub>>>k<sub>3</sub>,k<sub>6</sub>:it is hard to predict the value of k<sub>3</sub> and k<sub>8</sub>, yet we suppose both of them is much smaller than k<sub>10</sub> because SdpI is a kind of membrane protein inherently, and rarely exists as free protein</li><br />
<li>The primary values of all the six variables are very small or strictly zero. We expect it as the most logical initial status. If the primary value of any variable is relatively large, the suicide mechanism may not run normally</li><br />
</ol><br />
</br></br><br />
<h2>Stimulation and discussion</h2><br />
<br />
Simple and rough as the above model is, it does theoretically sound. To test the validity of this model, we first tried to get analytic solution of the ODE set. If this analytic solution exists, we could further investigate the interaction among those variables, and draw some phase planes to get accurate and mathematically perfect description of this model. Unfortunately but expectedly, the existence of analytic solution was negated by MATLAB, and we had to assume groups of values for these constants in advance and analyze the arithmetic solutions instead. These arithmetic solutions not only justified this mechanism is effective enough to commit cell suicide but also indicated some unexpected, or even weird results that beyond our wildest imagination. There are two possibility account for the unexpected results: our model is too rough to include some assignable factor; or there are some implicit but objective limitation inside model, which may be substantiate by later experiments or papers.</br><br />
When we explored the arithmetic solutions of this ODE set, we received nearly one hundred warnings from MATLAB and for many times our most powerful computer ran out of its 8GB memory, but sometimes we can receive the solution within seconds. We had adjusted our parameters for several times before we got our first solution. Here is the values of parameters for this group, and the graph of arithmetic solutions is also given:</br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>50</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/2/26/Suicide1.png"></br></br><br />
At the first glance this graph seemed fine. Initially the concentration of SdpC decreased slightly due to the capturing of SdpI and the repression of float SdpR, but gradually the positive feedback loop works, and C<sub>f</sub> increases rapidly. But when we turned our attention to the curves of other parameters, things seemed not so perfect:</br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/c/cf/Suicide2.png"></br></br><br />
The curve of I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub> contradicted our common sense severely. First, I<sub>m</sub>>C<sub>i</sub>>R<sub>i</sub> is expected to be tenable all the time, which precludes the intersects among the three curves; Second, there is no mechanism in this system that could decrease their concentration, and all of them are increasing function; Third and most serious, never will them be negative, as they represent the concentration of real substances.<br />
<br />
Then we adjusted the parameters slightly for several times. To eliminate those absurd curves, we reconsidered some assumptions.<br />
Here we listed another representative group of parament values and relative graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/7/7d/Suicide3.png/800px-Suicide3.png"></br></br><br />
In this group, we gave up one former assumption and set k<sub>2</sub> equal to k<sub>9</sub>. We also gave positive values to I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub>, which were considered to be zero at first. And by groups of stimulations we realized the value of k<sub>2</sub> does matter, as the derivative of C<sub>f</sub> only increased slightly as k<sub>2</sub> lowers, and the positive values failed to avoid the weird phenomenon in the latter three curves.<br />
We also found that however we adjusted the primary value of I<sub>f</sub> and other parameters, If dropped into approximately zero extremely rapidly at the initial stage and remained balanced, which might account for why the derivatives of the latter curves were abnormally negative. Thus we modified another assumption and increased k<sub>7</sub>. Here is another group of values and corresponding graph:</br></br><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/1/15/Suicide4.png/800px-Suicide4.png" style="width:600; height:400;"></br></br><br />
Although the derivative of Im is not seriously positive constantly, the three latter curves seemed much more reasonable. Hence, we extrapolated although SdpI and SdpR share the same promoter, the expression of SdpI must much faster than SdpR to ensure successful “suicide.” Additionally, the increase of k<sub>7</sub> also represses SdpC, and hence the copy number of SdpC must be larger.<br />
We kept all other parameters constant and gradually augmented k<sub>0</sub>. The larger k<sub>0</sub>, the more perfect the curve seemed, and here are the values table and graph where k<sub>0</sub> equals 400, 80 times larger than k<sub>4</sub>.</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>400</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/3/33/Suicide5.png/800px-Suicide5.png"></br></br><br />
Take the curve of C<sub>f</sub> and R<sub>f</sub> separately, the curves seemed more perfect:</br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/9/9f/Suicide6.png"></br></br><br />
<br />
In wild bacteria who are unable to produced SdpC, naturally k<sub>0</sub> equals zero. We expected C<sub>f</sub> would decreased gradually and finally approximate zero, and here are the corresponding table and graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>30</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/1/19/Suicide7.png"></br></br><br />
Wired but not surprising, there were intersects among the latter three curve, and C<sub>f</sub> decreases continually when it is negative. We continued to try groups of these parameters, and this is the best one where we increased the primary concentration of SdpC and the normal expression rate of SdpI.<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/8/8e/Suicide8.png"></br></br><br />
The curve of C<sub>f</sub> and R<sub>f</sub> alone:<br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/e/eb/Suicide9.png"></br></br><br />
In spite of minimal abnormal phenomenon (C<sub>f</sub> was negative in later stage), this graph roughly testified that in wild bacterial the concentration of float SdpC will drop to nearly zero quickly.<br />
In sum, the ODE model of singular cells indicate following results:</br><br />
<ol><br />
<li>The character of free SdpC is most affected by k<sub>0</sub>, if the copy number of SdpC is large enough, it is theoretically reasonable to commit suicide;</li><br />
<li>The influence of the value of Imax and k<sub>2</sub> is much limited;</li><br />
<li>The amount of free SdpI is always near zero;</li><br />
<li>SdpC will not increase limitlessly however we transform parameters;</li><br />
<li>To ensure the success of suicide, it is required k<sub>0</sub>>>k<sub>4</sub>>>k<sub>7</sub>;</li><br />
The last conclusion was our biggest windfall, and we have verified the validity of this suicide mechanism in math. On the one hand, if further experiments proven #4 engineered bacteria will kill both siblings and themselves, it is highly like that the expression rate SdpI is much larger than SdpR even if they share the same promoter; on the other hand, if #4 engineered bacteria are not able to commit suicide, we can try to boost the expression of SdpI to adjust the bacteria.<br />
<br />
<br />
<h1>Discussion on colonies</h1><br />
<br />
In reality, the engineered bacteria aims at killing its siblings instead of itself, and at first almost all toxin SdpC will be secreted outside the bacteria. Assume the diffusion of toxin among cells comply with diffusion law, that is, the diffusion rate is proportionate with the gradient of concentration. Further assume the death concentration of SdpC is same to all bacteria expect those who contain this locus, the average life expectancy is bacteria will hinge on the rate and distribution of engineered bacteria, and the distribution of life expectancy of bacteria is similar to that of average free path of gas molecules.<br />
As long the coefficient of diffusion is large enough, any engineered bacterias, no matter how few, is adequate to devastate the whole colony. Alike to the average free path of thin gas, the average suicide time of the whole reporter system is inversely proportional with the square root of the rate of engineer bacteria containing this locus.<br />
<br />
<br />
<h1>References</h1><br />
Parallel pathways of repression and antirepression governing the transition to stationary phase in Bacillus subtilis<br />
AV Banse, A Chastanet, L Rahn-Lee…,PNAS ,2008 </br><br />
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<h1>Introduction</h1><br />
To be more user-friendly, 4# circuit contains a reporting system. After melting in water, the spores germinate and express blue pigment protein to report the best using time.</br><br />
Meanwhile, 4# circuit could also ensure biosafety. Because other circuit do not have self-killing device, 4# engineering bacterial should kill all the bacterial after using.</br><br />
<br />
</div><br />
<br />
<div><br />
<h1>Designing of the suicide system</h1><br />
We design a circuit of killing switch based on its endogenous genetic system.</br><br />
In B.subtilis, when it comes to the stationary phase, the environmental pressure increases and nutrition becomes limited, so B begin to produce spores. Now the community will be divided into two different parts. One of them are trying to kill others to get enough nutrient , delaying the production of spores and achieving a competitive advantage. Killing is mediated by the exported toxic protein SdpC. SdpI will appear on the membrane surface to avoid itself from being damaged. SdpI could bind free SdpC and autopressor SdpR, to remove SdpR’s inhibition against I and R, to produce more SdpI to offset SdpC, finally guaranteeing the subgroup alive, thereby delaying the spores production.</br><br />
<img src="https://static.igem.org/mediawiki/2013/2/2b/Reporter_3.png" width="500" height="350"><br />
We transfer SdpC which is fused by promoter SdpI/R into high copy plasmids in order to damage the balance of the system, thereby killing whole colony. When SdpC appears, SdpI on the membrane will bind free SdpC and adsorb SdpR to cease its inhibition against SdpI P/R, trying to produce more SdpI. At the same time, it will activate the promoter SdpR/I in our circuits and generate more SdpC.The system would fall into an infinite loop, and according to our modeling ,the amount of SdpC increases beyond the ability of SdpI.Thus,the cells with protection mechanism will crack and die because of too much SdpC. All above forms the killing device.<br />
We Also designed a test circuit,which contains promotor grac and sdpABC only,aiming to determine the ability of SdpC.</br><br />
<br />
<div></br></br><br />
There are both positive and negative feedback loops in this process. On the one hand, SdpI is unable to sequestrate the autorepressor, SdpR, until it captures the toxin, SdpC. The accumulation of SdpC will thus facilitate SdpI to capture more SdpR and thereby relieve the repression of SdpR, stimulating the expression of itself. This is the positive feedback loop which leads to the increasing accumulation of SdpC and finally the death of the bacteria. On the other hand, the removal of SdpR also enhance the expression of SdpI and accelerate the sequestration of SdpC, which forms a negative feedback loop whose effects contradict the positive feedback loop. However, since the copy number of SdpC is much higher, it is believed that the positive loop is strong enough to outweigh the negative one, which guarantees this mechanism will finally leads to collapse instead of equilibrium.<br />
</div><br />
<div><br />
<h1>The ODE model of singular cells</h1><br />
There is no denying fact that the essential goal of engineered bacterias who carry this so called “suicide” locus itself is to kill their siblings rather than themselves to ensure the survival of themselves. Surly they can kill their siblings, but can they finally eliminate themselves, as we expects? The trivial experiment protocol and huge uncertainty had put off our experiment, and as expected, we failed to achieve the construction of complete reporter system in our laboratory. Fortunately, we could resort to mathematical models to verify the validity of this locus theoretically. <br />
There are six independent variables in individual cells, and the theoretically if the initial conditions are fixed, all of them will be the univariate functions of time. The following table illustrates the mark and meaning of each variable:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub></td><br />
<td >Mole number of free SdpI in cytoplasm</a>.</td><br />
</tr><br />
<tr><br />
<td >I<sub>m</sub> </td><br />
<td >Mole number of SdpI in the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >C<sub>f</sub></td><br />
<td >Mole number of free SdpC in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >C<sub>i</sub></td><br />
<td >Mole number of SdpC captured by SdpI.</td><br />
</tr><br />
<tr><br />
<td >R<sub>f</sub></td><br />
<td >Mole number of free SdpR in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >R<sub>i</sub> </td><br />
<td >Mole number of SdpR captured by SdpI</td><br />
</tr><br />
</table><br />
</br></br><br />
To construct reasonable ordinary differential equation (ODE) model to describe and predict the operation of the suicide system, we followed the law of mass action, one basic law of chemistry and biology.</br><br />
Taken as a statement about kinetics, the law states that the rate of an elementary reaction (a reaction that proceeds through only one transition state, which is one mechanistic step) is proportional to the product of the concentrations of the participating molecules. In modern chemistry this is derived using statistical mechanics. Despite the complicated chemical reactions involved in the process of transcription and translation, it is common and logically sound to view the expression of one particular gene as an elementary reaction and assume the repression effects of the protein itself encodes and the repressor are both linear.</br><br />
According to the law of mass action, we got six independent differential equation of the variables:</br><br />
<img src="https://static.igem.org/mediawiki/2013/e/e8/For1%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/2/23/For2%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/4/48/For3%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/7/71/For4%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/3/31/For5%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/c/cb/For6%281%29.png"></br></br></br><br />
The following table explain the constants in the above ODE groups:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub> </td><br />
<td >The maximal number of SdpI than can be fixed on the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >k<sub>0</sub></td><br />
<td >Constant describes the normal expression rate of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>1</sub> </td><br />
<td >Constant describes the self-repression effects of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>2</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpC.</a></td><br />
</tr><br />
<tr><br />
<td >k<sub>3</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>4</sub> </td><br />
<td >Constant describes the normal expression rate of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>5</sub> </td><br />
<td >Constant describes the self-repression effects of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>6</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>7</sub> </td><br />
<td >Constant describes the normal expression rate of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>8</sub> </td><br />
<td >Constant describes the self-repression effects of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>9</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>10</sub> </td><br />
<td >Constant describes the rate of SdpI binding to the cell membrane</td><br />
</tr><br />
</table><br />
<br />
<br />
<br />
<br />
</div><br />
<div><br />
<h2>Discussions on the constants</h2><br />
All the constants given above is steady and theoretically measurable when all the conditions are constant. For example, we could measure k<sub>0</sub> by constructing a new engineered bacteria, which contains the gene encoding SdpC and marker gene alone and observing the influence of the concentration of SdpC on its expression. Yet any modification on genome is notoriously time-consuming, which inhibited us from measuring them in person. We also looked up oceans of papers to confer their approximate ranges, but almost all papers are too fragmental to afford any valid information. Therefore, we decided to assume all these constant according to our limited information and make a qualitative analysis instead of quantifiable analysis. All units and dimensions were temporarily ignored. In other words, our model aims at justifying the validity of this suicide mechanism rather than predicting the exact time or any other parameters of the system.<br />
<br />
Despite the fact that we have hardly any accurate data on these constants, there are some limitations that we extrapolated from known information before we further explore this model:</br><br />
<ol><br />
<li>k<sub>0</sub>>>k<sub>4</sub>≈k<sub>7</sub>: k<sub>0</sub>,k<sub>4</sub> and k<sub>7</sub> represent the normal expression rate of SdpC, SdpR and SdpC separately, and the copy number of SdpC is much larger than that of SdpR and SdpI, whereas the value of the latter two is approximately equal;</li><br />
<li>k<sub>2</sub>>>k<sub>9</sub>: the existence of free SdpR represses the expression of both SdpI and SdpC, and similarly, since the copy number of SdpC is much higher, we expected the repression effect was stronger accordingly;</li><br />
<li>k<sub>10</sub>>>k<sub>3</sub>,k<sub>6</sub>:it is hard to predict the value of k<sub>3</sub> and k<sub>8</sub>, yet we suppose both of them is much smaller than k<sub>10</sub> because SdpI is a kind of membrane protein inherently, and rarely exists as free protein</li><br />
<li>The primary values of all the six variables are very small or strictly zero. We expect it as the most logical initial status. If the primary value of any variable is relatively large, the suicide mechanism may not run normally</li><br />
</ol><br />
</br></br><br />
<h2>Stimulation and discussion</h2><br />
<br />
Simple and rough as the above model is, it does theoretically sound. To test the validity of this model, we first tried to get analytic solution of the ODE set. If this analytic solution exists, we could further investigate the interaction among those variables, and draw some phase planes to get accurate and mathematically perfect description of this model. Unfortunately but expectedly, the existence of analytic solution was negated by MATLAB, and we had to assume groups of values for these constants in advance and analyze the arithmetic solutions instead. These arithmetic solutions not only justified this mechanism is effective enough to commit cell suicide but also indicated some unexpected, or even weird results that beyond our wildest imagination. There are two possibility account for the unexpected results: our model is too rough to include some assignable factor; or there are some implicit but objective limitation inside model, which may be substantiate by later experiments or papers.</br><br />
When we explored the arithmetic solutions of this ODE set, we received nearly one hundred warnings from MATLAB and for many times our most powerful computer ran out of its 8GB memory, but sometimes we can receive the solution within seconds. We had adjusted our parameters for several times before we got our first solution. Here is the values of parameters for this group, and the graph of arithmetic solutions is also given:</br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>50</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/2/26/Suicide1.png"></br></br><br />
At the first glance this graph seemed fine. Initially the concentration of SdpC decreased slightly due to the capturing of SdpI and the repression of float SdpR, but gradually the positive feedback loop works, and C<sub>f</sub> increases rapidly. But when we turned our attention to the curves of other parameters, things seemed not so perfect:</br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/c/cf/Suicide2.png"></br></br><br />
The curve of I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub> contradicted our common sense severely. First, I<sub>m</sub>>C<sub>i</sub>>R<sub>i</sub> is expected to be tenable all the time, which precludes the intersects among the three curves; Second, there is no mechanism in this system that could decrease their concentration, and all of them are increasing function; Third and most serious, never will them be negative, as they represent the concentration of real substances.<br />
<br />
Then we adjusted the parameters slightly for several times. To eliminate those absurd curves, we reconsidered some assumptions.<br />
Here we listed another representative group of parament values and relative graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/7/7d/Suicide3.png/800px-Suicide3.png"></br></br><br />
In this group, we gave up one former assumption and set k<sub>2</sub> equal to k<sub>9</sub>. We also gave positive values to I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub>, which were considered to be zero at first. And by groups of stimulations we realized the value of k<sub>2</sub> does matter, as the derivative of C<sub>f</sub> only increased slightly as k<sub>2</sub> lowers, and the positive values failed to avoid the weird phenomenon in the latter three curves.<br />
We also found that however we adjusted the primary value of I<sub>f</sub> and other parameters, If dropped into approximately zero extremely rapidly at the initial stage and remained balanced, which might account for why the derivatives of the latter curves were abnormally negative. Thus we modified another assumption and increased k<sub>7</sub>. Here is another group of values and corresponding graph:</br></br><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/1/15/Suicide4.png/800px-Suicide4.png" style="width:600; height:400;"></br></br><br />
Although the derivative of Im is not seriously positive constantly, the three latter curves seemed much more reasonable. Hence, we extrapolated although SdpI and SdpR share the same promoter, the expression of SdpI must much faster than SdpR to ensure successful “suicide.” Additionally, the increase of k<sub>7</sub> also represses SdpC, and hence the copy number of SdpC must be larger.<br />
We kept all other parameters constant and gradually augmented k<sub>0</sub>. The larger k<sub>0</sub>, the more perfect the curve seemed, and here are the values table and graph where k<sub>0</sub> equals 400, 80 times larger than k<sub>4</sub>.</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>400</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/3/33/Suicide5.png/800px-Suicide5.png"></br></br><br />
Take the curve of C<sub>f</sub> and R<sub>f</sub> separately, the curves seemed more perfect:</br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/9/9f/Suicide6.png"></br></br><br />
<br />
In wild bacteria who are unable to produced SdpC, naturally k<sub>0</sub> equals zero. We expected C<sub>f</sub> would decreased gradually and finally approximate zero, and here are the corresponding table and graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>30</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/1/19/Suicide7.png"></br></br><br />
Wired but not surprising, there were intersects among the latter three curve, and C<sub>f</sub> decreases continually when it is negative. We continued to try groups of these parameters, and this is the best one where we increased the primary concentration of SdpC and the normal expression rate of SdpI.<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/8/8e/Suicide8.png"></br></br><br />
The curve of C<sub>f</sub> and R<sub>f</sub> alone:<br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/e/eb/Suicide9.png"></br></br><br />
In spite of minimal abnormal phenomenon (C<sub>f</sub> was negative in later stage), this graph roughly testified that in wild bacterial the concentration of float SdpC will drop to nearly zero quickly.<br />
In sum, the ODE model of singular cells indicate following results:</br><br />
<ol><br />
<li>The character of free SdpC is most affected by k<sub>0</sub>, if the copy number of SdpC is large enough, it is theoretically reasonable to commit suicide;</li><br />
<li>The influence of the value of Imax and k<sub>2</sub> is much limited;</li><br />
<li>The amount of free SdpI is always near zero;</li><br />
<li>SdpC will not increase limitlessly however we transform parameters;</li><br />
<li>To ensure the success of suicide, it is required k<sub>0</sub>>>k<sub>4</sub>>>k<sub>7</sub>;</li><br />
The last conclusion was our biggest windfall, and we have verified the validity of this suicide mechanism in math. On the one hand, if further experiments proven #4 engineered bacteria will kill both siblings and themselves, it is highly like that the expression rate SdpI is much larger than SdpR even if they share the same promoter; on the other hand, if #4 engineered bacteria are not able to commit suicide, we can try to boost the expression of SdpI to adjust the bacteria.<br />
<br />
<br />
<h1>Discussion on colonies</h1><br />
<br />
In reality, the engineered bacteria aims at killing its siblings instead of itself, and at first almost all toxin SdpC will be secreted outside the bacteria. Assume the diffusion of toxin among cells comply with diffusion law, that is, the diffusion rate is proportionate with the gradient of concentration. Further assume the death concentration of SdpC is same to all bacteria expect those who contain this locus, the average life expectancy is bacteria will hinge on the rate and distribution of engineered bacteria, and the distribution of life expectancy of bacteria is similar to that of average free path of gas molecules.<br />
As long the coefficient of diffusion is large enough, any engineered bacterias, no matter how few, is adequate to devastate the whole colony. Alike to the average free path of thin gas, the average suicide time of the whole reporter system is inversely proportional with the square root of the rate of engineer bacteria containing this locus.<br />
<br />
<br />
<h1>References</h1><br />
Parallel pathways of repression and antirepression governing the transition to stationary phase in Bacillus subtilis<br />
AV Banse, A Chastanet, L Rahn-Lee…,PNAS ,2008 </br><br />
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<h1>Introduction</h1><br />
To be more user-friendly, 4# circuit contains a reporting system. After melting in water, the spores germinate and express blue pigment protein to report the best using time.</br><br />
Meanwhile, 4# circuit could also ensure biosafety. Because other circuit do not have self-killing device, 4# engineering bacterial should kill all the bacterial after using.</br><br />
<br />
</div><br />
<br />
<div><br />
<h1>Designing of the suicide system</h1><br />
<p>We design a circuit of killing switch based on its endogenous genetic system.</br><br />
In B.subtilis, when it comes to the stationary phase, the environmental pressure increases and nutrition becomes limited, so B begin to produce spores. Now the community will be divided into two different parts. One of them are trying to kill others to get enough nutrient , delaying the production of spores and achieving a competitive advantage. Killing is mediated by the exported toxic protein SdpC. SdpI will appear on the membrane surface to avoid itself from being damaged. SdpI could bind free SdpC and autopressor SdpR, to remove SdpR’s inhibition against I and R, to produce more SdpI to offset SdpC, finally guaranteeing the subgroup alive, thereby delaying the spores production.</p><br />
<img src="https://static.igem.org/mediawiki/2013/2/2b/Reporter_3.png" width="500" height="350"><br />
<p>We transfer SdpC which is fused by promoter SdpI/R into high copy plasmids in order to damage the balance of the system, thereby killing whole colony. When SdpC appears, SdpI on the membrane will bind free SdpC and adsorb SdpR to cease its inhibition against SdpI P/R, trying to produce more SdpI. At the same time, it will activate the promoter SdpR/I in our circuits and generate more SdpC.The system would fall into an infinite loop, and according to our modeling ,the amount of SdpC increases beyond the ability of SdpI.Thus,the cells with protection mechanism will crack and die because of too much SdpC. All above forms the killing device.<br />
We Also designed a test circuit,which contains promotor grac and sdpABC only,aiming to determine the ability of SdpC.</p><br />
</div><br />
<div><br />
<p><br />
There are both positive and negative feedback loops in this process. On the one hand, SdpI is unable to sequestrate the autorepressor, SdpR, until it captures the toxin, SdpC. The accumulation of SdpC will thus facilitate SdpI to capture more SdpR and thereby relieve the repression of SdpR, stimulating the expression of itself. This is the positive feedback loop which leads to the increasing accumulation of SdpC and finally the death of the bacteria. On the other hand, the removal of SdpR also enhance the expression of SdpI and accelerate the sequestration of SdpC, which forms a negative feedback loop whose effects contradict the positive feedback loop. However, since the copy number of SdpC is much higher, it is believed that the positive loop is strong enough to outweigh the negative one, which guarantees this mechanism will finally leads to collapse instead of equilibrium.</p><br />
</div><br />
<div><br />
<h1>The ODE model of singular cells</h1><br />
<p>There is no denying fact that the essential goal of engineered bacterias who carry this so called “suicide” locus itself is to kill their siblings rather than themselves to ensure the survival of themselves. Surly they can kill their siblings, but can they finally eliminate themselves, as we expects? The trivial experiment protocol and huge uncertainty had put off our experiment, and as expected, we failed to achieve the construction of complete reporter system in our laboratory. Fortunately, we could resort to mathematical models to verify the validity of this locus theoretically.<br> <br />
There are six independent variables in individual cells, and the theoretically if the initial conditions are fixed, all of them will be the univariate functions of time. The following table illustrates the mark and meaning of each variable:<p><br />
<br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub></td><br />
<td >Mole number of free SdpI in cytoplasm</a>.</td><br />
</tr><br />
<tr><br />
<td >I<sub>m</sub> </td><br />
<td >Mole number of SdpI in the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >C<sub>f</sub></td><br />
<td >Mole number of free SdpC in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >C<sub>i</sub></td><br />
<td >Mole number of SdpC captured by SdpI.</td><br />
</tr><br />
<tr><br />
<td >R<sub>f</sub></td><br />
<td >Mole number of free SdpR in cytoplasm.</td><br />
</tr><br />
<tr><br />
<td >R<sub>i</sub> </td><br />
<td >Mole number of SdpR captured by SdpI</td><br />
</tr><br />
</table><br />
<p><br />
To construct reasonable ordinary differential equation (ODE) model to describe and predict the operation of the suicide system, we followed the law of mass action, one basic law of chemistry and biology.</br><br />
Taken as a statement about kinetics, the law states that the rate of an elementary reaction (a reaction that proceeds through only one transition state, which is one mechanistic step) is proportional to the product of the concentrations of the participating molecules. In modern chemistry this is derived using statistical mechanics. Despite the complicated chemical reactions involved in the process of transcription and translation, it is common and logically sound to view the expression of one particular gene as an elementary reaction and assume the repression effects of the protein itself encodes and the repressor are both linear.<p><br />
<p>According to the law of mass action, we got six independent differential equation of the variables:</P><br />
<img src="https://static.igem.org/mediawiki/2013/e/e8/For1%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/2/23/For2%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/4/48/For3%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/7/71/For4%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/3/31/For5%281%29.png"></br></br></br><br />
<img src="https://static.igem.org/mediawiki/2013/c/cb/For6%281%29.png"></br></br></br><br />
<p>The following table explain the constants in the above ODE groups:</p><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td >Mark</td><br />
<td >Meaning</td><br />
</tr><br />
<tr><br />
<td >I<sub>max</sub> </td><br />
<td >The maximal number of SdpI than can be fixed on the cell membrane.</td><br />
</tr><br />
<tr><br />
<td >k<sub>0</sub></td><br />
<td >Constant describes the normal expression rate of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>1</sub> </td><br />
<td >Constant describes the self-repression effects of SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>2</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpC.</a></td><br />
</tr><br />
<tr><br />
<td >k<sub>3</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpC</td><br />
</tr><br />
<tr><br />
<td >k<sub>4</sub> </td><br />
<td >Constant describes the normal expression rate of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>5</sub> </td><br />
<td >Constant describes the self-repression effects of SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>6</sub> </td><br />
<td >Constant describes the rate of SdpI capturing SdpR</td><br />
</tr><br />
<tr><br />
<td >k<sub>7</sub> </td><br />
<td >Constant describes the normal expression rate of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>8</sub> </td><br />
<td >Constant describes the self-repression effects of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>9</sub> </td><br />
<td >Constant describes the repression of SdpR on the expression of SdpI</td><br />
</tr><br />
<tr><br />
<td >k<sub>10</sub> </td><br />
<td >Constant describes the rate of SdpI binding to the cell membrane</td><br />
</tr><br />
</table><br />
</div><br />
<br />
<div><br />
<h2>Discussions on the constants</h2><br />
<p>All the constants given above is steady and theoretically measurable when all the conditions are constant. For example, we could measure k<sub>0</sub> by constructing a new engineered bacteria, which contains the gene encoding SdpC and marker gene alone and observing the influence of the concentration of SdpC on its expression. Yet any modification on genome is notoriously time-consuming, which inhibited us from measuring them in person. We also looked up oceans of papers to confer their approximate ranges, but almost all papers are too fragmental to afford any valid information. Therefore, we decided to assume all these constant according to our limited information and make a qualitative analysis instead of quantifiable analysis. All units and dimensions were temporarily ignored. In other words, our model aims at justifying the validity of this suicide mechanism rather than predicting the exact time or any other parameters of the system.<br><br />
<br />
Despite the fact that we have hardly any accurate data on these constants, there are some limitations that we extrapolated from known information before we further explore this model:</p><br />
<ol><br />
<li>k<sub>0</sub>>>k<sub>4</sub>≈k<sub>7</sub>: k<sub>0</sub>,k<sub>4</sub> and k<sub>7</sub> represent the normal expression rate of SdpC, SdpR and SdpC separately, and the copy number of SdpC is much larger than that of SdpR and SdpI, whereas the value of the latter two is approximately equal;</li><br />
<li>k<sub>2</sub>>>k<sub>9</sub>: the existence of free SdpR represses the expression of both SdpI and SdpC, and similarly, since the copy number of SdpC is much higher, we expected the repression effect was stronger accordingly;</li><br />
<li>k<sub>10</sub>>>k<sub>3</sub>,k<sub>6</sub>:it is hard to predict the value of k<sub>3</sub> and k<sub>8</sub>, yet we suppose both of them is much smaller than k<sub>10</sub> because SdpI is a kind of membrane protein inherently, and rarely exists as free protein</li><br />
<li>The primary values of all the six variables are very small or strictly zero. We expect it as the most logical initial status. If the primary value of any variable is relatively large, the suicide mechanism may not run normally</li><br />
</ol><br />
</br></br><br />
<h2>Stimulation and discussion</h2><br />
<br />
<p>Simple and rough as the above model is, it does theoretically sound. To test the validity of this model, we first tried to get analytic solution of the ODE set. If this analytic solution exists, we could further investigate the interaction among those variables, and draw some phase planes to get accurate and mathematically perfect description of this model. Unfortunately but expectedly, the existence of analytic solution was negated by MATLAB, and we had to assume groups of values for these constants in advance and analyze the arithmetic solutions instead. These arithmetic solutions not only justified this mechanism is effective enough to commit cell suicide but also indicated some unexpected, or even weird results that beyond our wildest imagination. There are two possibility account for the unexpected results: our model is too rough to include some assignable factor; or there are some implicit but objective limitation inside model, which may be substantiate by later experiments or papers.</br><br />
When we explored the arithmetic solutions of this ODE set, we received nearly one hundred warnings from MATLAB and for many times our most powerful computer ran out of its 8GB memory, but sometimes we can receive the solution within seconds. We had adjusted our parameters for several times before we got our first solution. Here is the values of parameters for this group, and the graph of arithmetic solutions is also given:</p><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>50</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/2/26/Suicide1.png"></br></br><br />
At the first glance this graph seemed fine. Initially the concentration of SdpC decreased slightly due to the capturing of SdpI and the repression of float SdpR, but gradually the positive feedback loop works, and C<sub>f</sub> increases rapidly. But when we turned our attention to the curves of other parameters, things seemed not so perfect:</br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/c/cf/Suicide2.png"></br></br><br />
The curve of I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub> contradicted our common sense severely. First, I<sub>m</sub>>C<sub>i</sub>>R<sub>i</sub> is expected to be tenable all the time, which precludes the intersects among the three curves; Second, there is no mechanism in this system that could decrease their concentration, and all of them are increasing function; Third and most serious, never will them be negative, as they represent the concentration of real substances.<br />
<br />
Then we adjusted the parameters slightly for several times. To eliminate those absurd curves, we reconsidered some assumptions.<br />
Here we listed another representative group of parament values and relative graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/7/7d/Suicide3.png/800px-Suicide3.png"></br></br><br />
In this group, we gave up one former assumption and set k<sub>2</sub> equal to k<sub>9</sub>. We also gave positive values to I<sub>m</sub>, C<sub>i</sub> and R<sub>i</sub>, which were considered to be zero at first. And by groups of stimulations we realized the value of k<sub>2</sub> does matter, as the derivative of C<sub>f</sub> only increased slightly as k<sub>2</sub> lowers, and the positive values failed to avoid the weird phenomenon in the latter three curves.<br />
We also found that however we adjusted the primary value of I<sub>f</sub> and other parameters, If dropped into approximately zero extremely rapidly at the initial stage and remained balanced, which might account for why the derivatives of the latter curves were abnormally negative. Thus we modified another assumption and increased k<sub>7</sub>. Here is another group of values and corresponding graph:</br></br><br />
<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>100</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/1/15/Suicide4.png/800px-Suicide4.png" style="width:600; height:400;"></br></br><br />
Although the derivative of Im is not seriously positive constantly, the three latter curves seemed much more reasonable. Hence, we extrapolated although SdpI and SdpR share the same promoter, the expression of SdpI must much faster than SdpR to ensure successful “suicide.” Additionally, the increase of k<sub>7</sub> also represses SdpC, and hence the copy number of SdpC must be larger.<br />
We kept all other parameters constant and gradually augmented k<sub>0</sub>. The larger k<sub>0</sub>, the more perfect the curve seemed, and here are the values table and graph where k<sub>0</sub> equals 400, 80 times larger than k<sub>4</sub>.</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>400</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>5</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/thumb/3/33/Suicide5.png/800px-Suicide5.png"></br></br><br />
Take the curve of C<sub>f</sub> and R<sub>f</sub> separately, the curves seemed more perfect:</br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/9/9f/Suicide6.png"></br></br><br />
<br />
In wild bacteria who are unable to produced SdpC, naturally k<sub>0</sub> equals zero. We expected C<sub>f</sub> would decreased gradually and finally approximate zero, and here are the corresponding table and graph:</br></br><br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>30</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/1/19/Suicide7.png"></br></br><br />
Wired but not surprising, there were intersects among the latter three curve, and C<sub>f</sub> decreases continually when it is negative. We continued to try groups of these parameters, and this is the best one where we increased the primary concentration of SdpC and the normal expression rate of SdpI.<br />
<table border="1" align="center" frame="box"><br />
<tr><br />
<td>k<sub>0</sub></td><br />
<td>k<sub>1</sub></td><br />
<td>k<sub>2</sub></td><br />
<td>k<sub>3</sub></td><br />
<td>k<sub>4</sub></td><br />
<td>k<sub>5</sub></td><br />
<td>k<sub>6</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>8</sub></td><br />
<td>k<sub>9</sub></td><br />
<td>k<sub>10</sub></td><br />
<td>I<sub>max</sub></td><br />
<td>C<sub>f0</sub></td><br />
<td>R<sub>f0</sub></td><br />
<td>I<sub>f0</sub></td><br />
<td>I<sub>m0</sub></td><br />
<td>C<sub>i0</sub></td><br />
<td>R<sub>i0</sub></td><br />
</tr><br />
<tr><br />
<td>0</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>5</td><br />
<td>20</td><br />
<td>500</td><br />
<td>8</td><br />
<td>5</td><br />
<td>1</td><br />
<td>5</td><br />
<td>3</td><br />
<td>2</td><br />
</tr><br />
</table></br></br><br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/8/8e/Suicide8.png"></br></br><br />
The curve of C<sub>f</sub> and R<sub>f</sub> alone:<br />
<img class="linegraph" src="https://static.igem.org/mediawiki/2013/e/eb/Suicide9.png"></br></br><br />
In spite of minimal abnormal phenomenon (C<sub>f</sub> was negative in later stage), this graph roughly testified that in wild bacterial the concentration of float SdpC will drop to nearly zero quickly.<br />
In sum, the ODE model of singular cells indicate following results:</br><br />
<ol><br />
<li>The character of free SdpC is most affected by k<sub>0</sub>, if the copy number of SdpC is large enough, it is theoretically reasonable to commit suicide;</li><br />
<li>The influence of the value of Imax and k<sub>2</sub> is much limited;</li><br />
<li>The amount of free SdpI is always near zero;</li><br />
<li>SdpC will not increase limitlessly however we transform parameters;</li><br />
<li>To ensure the success of suicide, it is required k<sub>0</sub>>>k<sub>4</sub>>>k<sub>7</sub>;</li><br />
The last conclusion was our biggest windfall, and we have verified the validity of this suicide mechanism in math. On the one hand, if further experiments proven #4 engineered bacteria will kill both siblings and themselves, it is highly like that the expression rate SdpI is much larger than SdpR even if they share the same promoter; on the other hand, if #4 engineered bacteria are not able to commit suicide, we can try to boost the expression of SdpI to adjust the bacteria.<br />
<br />
<br />
<h1>Discussion on colonies</h1><br />
<br />
In reality, the engineered bacteria aims at killing its siblings instead of itself, and at first almost all toxin SdpC will be secreted outside the bacteria. Assume the diffusion of toxin among cells comply with diffusion law, that is, the diffusion rate is proportionate with the gradient of concentration. Further assume the death concentration of SdpC is same to all bacteria expect those who contain this locus, the average life expectancy is bacteria will hinge on the rate and distribution of engineered bacteria, and the distribution of life expectancy of bacteria is similar to that of average free path of gas molecules.<br />
As long the coefficient of diffusion is large enough, any engineered bacterias, no matter how few, is adequate to devastate the whole colony. Alike to the average free path of thin gas, the average suicide time of the whole reporter system is inversely proportional with the square root of the rate of engineer bacteria containing this locus.<br />
<br />
<br />
<h1>References</h1><br />
Parallel pathways of repression and antirepression governing the transition to stationary phase in Bacillus subtilis<br />
AV Banse, A Chastanet, L Rahn-Lee…,PNAS ,2008 </br><br />
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<li><a href="https://igem.org/2013_Judging_Form?id=1074#iGEM_Medals">Achievements</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li><br />
</ul><br />
<div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" /><br />
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<br />
<div class="content" align="center"><br />
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<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></div></div><br />
<div class="conbar2"><br />
<div class="leftbar" align="left" style="margin-bottom:30px;"><br />
<br />
<br />
<div class="part-col-1"><br />
<h1>TD1, Transdermal peptide</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074000">BBa_K1074000</a></h2><br />
<p>TD1 is a short synthetic peptide(ACSSSPSKHCG) identified by in vivo phage display, facilitated efficient transdermal protein delivery through intact skin...</p><br />
</div><br />
<br />
<div class="part-col-1"><br />
<h1>Pgrac+RBS+SamyQ+TD1<br>+GFP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074006">BBa_K1074006</a></h2><br />
<p>This is the main circuit of our project to allow high expression of target protein(antigen,adjuvant).GFP can be substituted by various proteins...</p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>PHT43</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074001">BBa_K1074001</a></h2><br />
<p>PHT43 is a E.coli-B.subtilis shuttle vector allowing high-level expression of secreted proteins in B.subtilis... </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>HBsAg</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074002">BBa_K1074002</a></h2> <br />
<p>HBsAg is the surface antigen of the hepatitis B virus (HBV). It indicates current hepatitis B infection...</p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>Ag85b</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074003">BBa_K1074003</a></h2> <br />
<p>The antigen 85 proteins (FbpA, FbpB, FbpC) are responsible for the high affinity of mycobacteria for fibronectin... </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>Protective Antigen Domain 4</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074004">BBa_K1074004</a></h2> <br />
<p>Protective antigen (PA) is the central component of the three-part protein toxin secreted by Bacillus anthracis, the organism responsible for anthrax... </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>LTB</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074005">BBa_K1074005</a></h2> <br />
<p>This eltB gene encodes for the Heat-labile enterotox(LT) in certain virulent strains of E.coli...</p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Pctc+RBS+<br>amilCP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074007">BBa_K1074007</a></h2> <br />
<p>Promoter ctc(BBa_K143010) is a sigma factor B-dependent promoter in B. subtilis. activated by endogenous sigma factor B ...</p><br />
</div><br />
<div class="part-col-2"><br />
<h1>SdpA,SdpB,<br>SpbC</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074009">BBa_K1074009</a></h2> <br />
<p>Killing factor of Bacillus subtilis ,SpbC Induces the lysis of other B.subtilis cells that have not entered the sporulation pathway...</p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Pgrac+RBS+SDP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074010">BBa_K1074010</a></h2> <br />
<p>Spbc in SDP(sdpA,sdpB,spbC)(BBa_K1074009)operon is a killing factor of Bacillus subtilis...</p><br />
</div><br />
<div class="clear"></div><br />
<div class="part-col-2"><br />
<h1>PsdpRI+RBS<br>+SDP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074011">BBa_K1074011</a></h2> <br />
<p>We construct this gene circuit as our kill switch to kill the engineered Bacillus subtilis for the safety purpose...</p><br />
</div><br />
<div class="part-col-2"><br />
<br />
<br />
<br />
<h1>Pgrac promoter</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074012">BBa_K1074012</a></h2> <br />
<p>Prgac promoter(consisting of the groE promoter,the lacO operator and the gsiBSD sequence) allow induction by addition of ITPG... </p><br />
</div><br />
<br />
<br />
<br />
<div class="part-col-2"><br />
<h1>promoter sdpR/I</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074013">BBa_K1074013</a></h2> <br />
<p>This part is a sigma factor A-dependent promotor of the gene sdpR derived from the B.subtilis...</p><br />
</div><br />
<div class="part-col-2"><br />
<br />
<br />
<h1>SamyQ</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074014">BBa_K1074014</a></h2> <br />
<p>SamyQ is the signal peptide of the amyQ gene encoding an α-amylase in Bacillus subtilis WB800N...</p><br />
</div><br />
<br />
<br />
<br />
<div class="part-col-2"><br />
<h1>RBS for Pgrac</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074015">BBa_K1074015</a></h2> <br />
<p>An efficient RBS for promoter Pgrac(BBa_K1074012) in Bacillus subtilis.../p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>RBS for Pctc</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074016">BBa_K1074016</a></h2> <br />
<p>This is an efficient RBS for promoter Pctc(BBa_K143010) in Bacillus subtilis...</p><br />
</div><br />
<br />
<div class="clear"></div><br />
</div><br />
<div class="rightbar"><br />
<div class="port-sidebar-border"><h>Project</h></div><br />
<div class="clear"></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails">Project Details</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails/Background">Background</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails/Design">Design</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Basic Experiment</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results/FurthurWork">Furthur Work</a></div><br />
<div id="t1"><a class="active" href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></div><br />
</div></div></div> <br />
<br />
</body><br />
</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/PartsTeam:USTC CHINA/Parts2013-09-27T14:41:11Z<p>Surine: </p>
<hr />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">Notebook</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">Timeline</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></li><br />
</ul><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li><br />
</ul><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li><br />
</ul><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Members</a></li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=USTC_CHINA">Profile</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Attributions">Attributions</a></li><br />
<li><a href="https://igem.org/2013_Judging_Form?id=1074#iGEM_Medals">Achievements</a></li><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li><br />
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<div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" /><br />
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<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></div></div><br />
<div class="conbar2"><br />
<div class="leftbar" align="left" style="margin-bottom:30px;"><br />
<br />
<br />
<div class="part-col-1"><br />
<h1>TD1, Transdermal peptide</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074000">BBa_K1074000</a></h2><br />
<p>TD1 is a short synthetic peptide(ACSSSPSKHCG) identified by in vivo phage display, facilitated efficient transdermal protein delivery through intact skin...</p><br />
</div><br />
<br />
<div class="part-col-1"><br />
<h1>Pgrac+RBS+SamyQ+TD1<br>+GFP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074006">BBa_K1074006</a></h2><br />
<p>This is the main circuit of our project to allow high expression of target protein(antigen,adjuvant).GFP can be substituted by various proteins...</p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>PHT43</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074001">BBa_K1074001</a></h2><br />
<p>PHT43 is a E.coli-B.subtilis shuttle vector allowing high-level expression of secreted proteins in B.subtilis... </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>HBsAg</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074002">BBa_K1074002</a></h2> <br />
<p>HBsAg is the surface antigen of the hepatitis B virus (HBV). It indicates current hepatitis B infection...</p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>Ag85b</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074003">BBa_K1074003</a></h2> <br />
<p>The antigen 85 proteins (FbpA, FbpB, FbpC) are responsible for the high affinity of mycobacteria for fibronectin... </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>Protective Antigen Domain 4</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074004">BBa_K1074004</a></h2> <br />
<p>Protective antigen (PA) is the central component of the three-part protein toxin secreted by Bacillus anthracis, the organism responsible for anthrax... </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>LTB</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074005">BBa_K1074005</a></h2> <br />
<p>This eltB gene encodes for the Heat-labile enterotox(LT) in certain virulent strains of E.coli...</p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Pctc+RBS+amilCP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074007">BBa_K1074007</a></h2> <br />
<p>Promoter ctc(BBa_K143010) is a sigma factor B-dependent promoter in B. subtilis. activated by endogenous sigma factor B ...</p><br />
</div><br />
<div class="part-col-2"><br />
<h1>SdpA,SdpB,SpbC</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074009">BBa_K1074009</a></h2> <br />
<p>Killing factor of Bacillus subtilis ,SpbC Induces the lysis of other B.subtilis cells that have not entered the sporulation pathway...</p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Pgrac+RBS+SDP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074010">BBa_K1074010</a></h2> <br />
<p>Spbc in SDP(sdpA,sdpB,spbC)(BBa_K1074009)operon is a killing factor of Bacillus subtilis...</p><br />
</div><br />
<div class="clear"></div><br />
<div class="part-col-2"><br />
<h1>PsdpRI+RBS+SDP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074011">BBa_K1074011</a></h2> <br />
<p>We construct this gene circuit as our kill switch to kill the engineered Bacillus subtilis for the safety purpose...</p><br />
</div><br />
<div class="part-col-2"><br />
<br />
<br />
<br />
<h1>Pgrac promoter</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074012">BBa_K1074012</a></h2> <br />
<p>Prgac promoter(consisting of the groE promoter,the lacO operator and the gsiBSD sequence) allow induction by addition of ITPG... </p><br />
</div><br />
<br />
<br />
<br />
<div class="part-col-2"><br />
<h1>promoter sdpR/I</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074013">BBa_K1074013</a></h2> <br />
<p>This part is a sigma factor A-dependent promotor of the gene sdpR derived from the B.subtilis...</p><br />
</div><br />
<div class="part-col-2"><br />
<br />
<br />
<h1>SamyQ</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074014">BBa_K1074014</a></h2> <br />
<p>SamyQ is the signal peptide of the amyQ gene encoding an α-amylase in Bacillus subtilis WB800N...</p><br />
</div><br />
<br />
<br />
<br />
<div class="part-col-2"><br />
<h1>RBS for Pgrac</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074015">BBa_K1074015</a></h2> <br />
<p>An efficient RBS for promoter Pgrac(BBa_K1074012) in Bacillus subtilis.../p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>RBS for Pctc</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074016">BBa_K1074016</a></h2> <br />
<p>This is an efficient RBS for promoter Pctc(BBa_K143010) in Bacillus subtilis...</p><br />
</div><br />
<br />
<div class="clear"></div><br />
</div><br />
<div class="rightbar"><br />
<div class="port-sidebar-border"><h>Project</h></div><br />
<div class="clear"></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails">Project Details</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails/Background">Background</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails/Design">Design</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Basic Experiment</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results/FurthurWork">Furthur Work</a></div><br />
<div id="t1"><a class="active" href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></div><br />
</div></div></div> <br />
<br />
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</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/PartsTeam:USTC CHINA/Parts2013-09-27T14:37:29Z<p>Surine: </p>
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></li><br />
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</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">Notebook</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">Timeline</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Members</a></li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=USTC_CHINA">Profile</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Attributions">Attributions</a></li><br />
<li><a href="https://igem.org/2013_Judging_Form?id=1074#iGEM_Medals">Achievements</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li><br />
</ul><br />
<div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" /><br />
</div><br />
</div><br />
</div><br />
<br />
<div class="content" align="center"><br />
<div class="conbar1"><br />
<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></div></div><br />
<div class="conbar2"><br />
<div class="leftbar" align="left" style="margin-bottom:30px;"><br />
<br />
<br />
<div class="part-col-1"><br />
<h1>TD1, Transdermal peptide</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074000">BBa_K1074000</a></h2><br />
<p>TD1 is a short synthetic peptide(ACSSSPSKHCG) identified by in vivo phage display, facilitated efficient transdermal protein delivery through intact skin. Studies suggested that the peptide creates a transient opening in the skin barrier to enable macromolecular material to reach systemic circulation.</p><br />
</div><br />
<br />
<div class="part-col-1"><br />
<h1>Pgrac+RBS+SamyQ+TD1<br>+GFP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074006">BBa_K1074006</a></h2><br />
<p>This is the main circuit of our project to allow high expression of target protein(antigen,adjuvant).GFP can be substituted by various proteins via a modular PCR or standard cut/ligation method.</p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>PHT43</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074001">BBa_K1074001</a></h2><br />
<p>PHT43 is a E.coli-B.subtilis shuttle vector allowing high-level expression of secreted proteins in B.subtilis. </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>HBsAg</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074002">BBa_K1074002</a></h2> <br />
<p>HBsAg is the surface antigen of the hepatitis B virus (HBV). It indicates current hepatitis B infection.</p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>Ag85b</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074003">BBa_K1074003</a></h2> <br />
<p>The antigen 85 proteins (FbpA, FbpB, FbpC) are responsible for the high affinity of mycobacteria for fibronectin, a large adhesive glycoprotein, which facilitates the attachment of M.tuberculosis to murine alveolar macrophages (AMs). </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>Protective Antigen Domain 4</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074004">BBa_K1074004</a></h2> <br />
<p>Protective antigen (PA) is the central component of the three-part protein toxin secreted by Bacillus anthracis, the organism responsible for anthrax. </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>LTB</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074005">BBa_K1074005</a></h2> <br />
<p>This eltB gene encodes for the Heat-labile enterotox(LT) in certain virulent strains of E.coli.</p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Pctc+RBS+amilCP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074007">BBa_K1074007</a></h2> <br />
<p>Promoter ctc(BBa_K143010) is a sigma factor B-dependent promoter in B. subtilis. activated by endogenous sigma factor B under mild stress( nutrient stress response or physical stress response). </p><br />
</div><br />
<div class="part-col-2"><br />
<h1>SdpA,SdpB,SpbC</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074009">BBa_K1074009</a></h2> <br />
<p>Killing factor of Bacillus subtilis ,SpbC Induces the lysis of other B.subtilis cells that have not entered the sporulation pathway, providing a source of nutrients to support sporulation.</p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Pgrac+RBS+SDP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074010">BBa_K1074010</a></h2> <br />
<p>Spbc in SDP(sdpA,sdpB,spbC)(BBa_K1074009)operon is a killing factor of Bacillus subtilis.</p><br />
</div><br />
<div class="clear"></div><br />
<div class="part-col-2"><br />
<h1>PsdpRI+RBS+SDP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074011">BBa_K1074011</a></h2> <br />
<p>We construct this gene circuit as our kill switch to kill the engineered Bacillus subtilis for the safety purpose.</p><br />
</div><br />
<div class="part-col-2"><br />
<br />
<br />
<br />
<h1>Pgrac promoter</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074012">BBa_K1074012</a></h2> <br />
<p>Prgac promoter(consisting of the groE promoter,the lacO operator and the gsiBSD sequence) allow induction by addition of ITPG. </p><br />
</div><br />
<br />
<br />
<br />
<div class="part-col-2"><br />
<h1>promoter sdpR/I</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074013">BBa_K1074013</a></h2> <br />
<p>This part is a sigma factor A-dependent promotor of the gene sdpR derived from the B.subtilis ,and it contains an operon for the signaling pathway of sdpc,a bacterial toxin of B.subtilis. </p><br />
</div><br />
<div class="part-col-2"><br />
<br />
<br />
<h1>SamyQ</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074014">BBa_K1074014</a></h2> <br />
<p>SamyQ is the signal peptide of the amyQ gene encoding an α-amylase in Bacillus subtilis WB800N. Fuse it with the recombinant proteins to obtain the secretion of recombinant proteins in Bacillus subtilis.</p><br />
</div><br />
<br />
<br />
<br />
<div class="part-col-2"><br />
<h1>RBS for Pgrac</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074015">BBa_K1074015</a></h2> <br />
<p>An efficient RBS for promoter Pgrac(BBa_K1074012) in Bacillus subtilis. We used it in all of our gene circuits containing promoter Pgrac. </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>RBS for Pctc</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074016">BBa_K1074016</a></h2> <br />
<p>This is an efficient RBS for promoter Pctc(BBa_K143010) in Bacillus subtilis. It's also an universal RBS for promoters in Bacillus subtilis. We also used it with the promoter PsdpRI(BBa_K1074013) in our project. </p><br />
</div><br />
<br />
<div class="clear"></div><br />
</div><br />
<div class="rightbar"><br />
<div class="port-sidebar-border"><h>Project</h></div><br />
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<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails">Project Details</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails/Background">Background</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails/Design">Design</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Basic Experiment</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results/FurthurWork">Furthur Work</a></div><br />
<div id="t1"><a class="active" href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></div><br />
</div></div></div> <br />
<br />
</body><br />
</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/PartsTeam:USTC CHINA/Parts2013-09-27T14:34:27Z<p>Surine: </p>
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li><br />
</ul><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li><br />
</ul><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Members</a></li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=USTC_CHINA">Profile</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Attributions">Attributions</a></li><br />
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</ul><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li><br />
</ul><br />
<div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" /><br />
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</div><br />
<br />
<div class="content" align="center"><br />
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<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></div></div><br />
<div class="conbar2"><br />
<div class="leftbar" align="left" style="margin-bottom:30px;"><br />
<br />
<br />
<div class="part-col-1"><br />
<h1>TD1, Transdermal peptide</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074000">BBa_K1074000</a></h2><br />
<p>TD1 is a short synthetic peptide(ACSSSPSKHCG) identified by in vivo phage display, facilitated efficient transdermal protein delivery through intact skin. Studies suggested that the peptide creates a transient opening in the skin barrier to enable macromolecular material to reach systemic circulation.</p><br />
</div><br />
<br />
<div class="part-col-1"><br />
<h1>Pgrac+RBS+SamyQ+TD1<br>+GFP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074006">BBa_K1074006</a></h2><br />
<p>This is the main circuit of our project to allow high expression of target protein(antigen,adjuvant).GFP can be substituted by various proteins via a modular PCR or standard cut/ligation method.</p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>PHT43</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074001">BBa_K1074001</a></h2><br />
<p>PHT43 is a E.coli-B.subtilis shuttle vector allowing high-level expression of secreted proteins in B.subtilis. </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>HBsAg</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074002">BBa_K1074002</a></h2> <br />
<p>HBsAg is the surface antigen of the hepatitis B virus (HBV). It indicates current hepatitis B infection.</p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>Ag85b</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074003">BBa_K1074003</a></h2> <br />
<p>The antigen 85 proteins (FbpA, FbpB, FbpC) are responsible for the high affinity of mycobacteria for fibronectin, a large adhesive glycoprotein, which facilitates the attachment of M.tuberculosis to murine alveolar macrophages (AMs). </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>Protective Antigen Domain 4</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074004">BBa_K1074004</a></h2> <br />
<p>Protective antigen (PA) is the central component of the three-part protein toxin secreted by Bacillus anthracis, the organism responsible for anthrax. </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>LTB</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074005">BBa_K1074005</a></h2> <br />
<p>This eltB gene encodes for the Heat-labile enterotox(LT) in certain virulent strains of E.coli.</p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Pctc+RBS+amilCP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074007">BBa_K1074007</a></h2> <br />
<p>Promoter ctc(BBa_K143010) is a sigma factor B-dependent promoter in B. subtilis. activated by endogenous sigma factor B under mild stress( nutrient stress response or physical stress response). </p><br />
</div><br />
<div class="part-col-2"><br />
<h1>SdpA,SdpB,SpbC</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074009">BBa_K1074009</a></h2> <br />
<p>Killing factor of Bacillus subtilis ,SpbC Induces the lysis of other B.subtilis cells that have not entered the sporulation pathway, providing a source of nutrients to support sporulation.</p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Pgrac+RBS+SDP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074010">BBa_K1074010</a></h2> <br />
<p>Spbc in SDP(sdpA,sdpB,spbC)(BBa_K1074009)operon is a killing factor of Bacillus subtilis.</p><br />
</div><br />
<div class="clear"></div><br />
<div class="part-col-2"><br />
<h1>PsdpRI+RBS+SDP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074011">BBa_K1074011</a></h2> <br />
<p>We construct this gene circuit as our kill switch to kill the engineered Bacillus subtilis for the safety purpose.Killing is mediated by the exported toxic protein SpbC. </p><br />
</div><br />
<div class="part-col-2"><br />
<br />
<br />
<br />
<h1>Pgrac promoter</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074012">BBa_K1074012</a></h2> <br />
<p>Prgac promoter(consisting of the groE promoter,the lacO operator and the gsiBSD sequence) allow induction by addition of ITPG. </p><br />
</div><br />
<br />
<br />
<br />
<div class="part-col-2"><br />
<h1>promoter sdpR/I</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074013">BBa_K1074013</a></h2> <br />
<p>This part is a sigma factor A-dependent promotor of the gene sdpR derived from the B.subtilis ,and it contains an operon for the signaling pathway of sdpc,a bacterial toxin of B.subtilis. </p><br />
</div><br />
<div class="part-col-2"><br />
<br />
<br />
<h1>SamyQ</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074014">BBa_K1074014</a></h2> <br />
<p>SamyQ is the signal peptide of the amyQ gene encoding an α-amylase in Bacillus subtilis WB800N. Fuse it with the recombinant proteins to obtain the secretion of recombinant proteins in Bacillus subtilis.</p><br />
</div><br />
<br />
<br />
<br />
<div class="part-col-2"><br />
<h1>RBS for Pgrac</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074015">BBa_K1074015</a></h2> <br />
<p>An efficient RBS for promoter Pgrac(BBa_K1074012) in Bacillus subtilis. We used it in all of our gene circuits containing promoter Pgrac. </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>RBS for Pctc</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074016">BBa_K1074016</a></h2> <br />
<p>This is an efficient RBS for promoter Pctc(BBa_K143010) in Bacillus subtilis. It's also an universal RBS for promoters in Bacillus subtilis. We also used it with the promoter PsdpRI(BBa_K1074013) in our project. </p><br />
</div><br />
<br />
<div class="clear"></div><br />
</div><br />
<div class="rightbar"><br />
<div class="port-sidebar-border"><h>Project</h></div><br />
<div class="clear"></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails">Project Details</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails/Background">Background</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails/Design">Design</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Basic Experiment</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results/FurthurWork">Furthur Work</a></div><br />
<div id="t1"><a class="active" href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></div><br />
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</body><br />
</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/PartsTeam:USTC CHINA/Parts2013-09-27T14:23:03Z<p>Surine: </p>
<hr />
<div>{{USTC-China/hidden}}<br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Members</a></li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=USTC_CHINA">Profile</a></li><br />
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<div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" /><br />
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<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></div></div><br />
<div class="conbar2"><br />
<div class="leftbar" align="left" style="margin-bottom:30px;"><br />
<br />
<br />
<div class="part-col-1"><br />
<h1>TD1, Transdermal peptide</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074000">BBa_K1074000</a></h2><br />
<p>TD1 is a short synthetic peptide(ACSSSPSKHCG) identified by in vivo phage display, facilitated efficient transdermal protein delivery through intact skin. Studies suggested that the peptide creates a transient opening in the skin barrier to enable macromolecular material to reach systemic circulation.</p><br />
</div><br />
<br />
<div class="part-col-1"><br />
<h1>Pgrac+RBS+SamyQ+TD1<br>+GFP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074006">BBa_K1074006</a></h2><br />
<p>This is the main circuit of our project to allow high expression of target protein(antigen,adjuvant).GFP can be substituted by various proteins via a modular PCR or standard cut/ligation method.</p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>PHT43</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074001">BBa_K1074001</a></h2><br />
<p>PHT43 is a E.coli-B.subtilis shuttle vector allowing high-level expression of secreted proteins in B.subtilis. </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>HBsAg</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074002">BBa_K1074002</a></h2> <br />
<p>HBsAg is the surface antigen of the hepatitis B virus (HBV). It indicates current hepatitis B infection.</p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>Ag85b</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074003">BBa_K1074003</a></h2> <br />
<p>The antigen 85 proteins (FbpA, FbpB, FbpC) are responsible for the high affinity of mycobacteria for fibronectin, a large adhesive glycoprotein, which facilitates the attachment of M.tuberculosis to murine alveolar macrophages (AMs). </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>Protective Antigen Domain 4</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074004">BBa_K1074004</a></h2> <br />
<p>Protective antigen (PA) is the central component of the three-part protein toxin secreted by Bacillus anthracis, the organism responsible for anthrax. </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>LTB</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074005">BBa_K1074005</a></h2> <br />
<p>This eltB gene encodes for the Heat-labile enterotox(LT) in certain virulent strains of E.coli.</p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Pctc+RBS+amilCP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074007">BBa_K1074007</a></h2> <br />
<p>Promoter ctc(BBa_K143010) is a sigma factor B-dependent promoter in B. subtilis. activated by endogenous sigma factor B under mild stress( nutrient stress response or physical stress response). </p><br />
</div><br />
<div class="part-col-2"><br />
<h1>SdpA,SdpB,SpbC</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074009">BBa_K1074009</a></h2> <br />
<p>Killing factor of Bacillus subtilis ,SpbC Induces the lysis of other B.subtilis cells that have not entered the sporulation pathway, providing a source of nutrients to support sporulation.</p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Pgrac+RBS+SDP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074010">BBa_K1074010</a></h2> <br />
<p>Spbc in SDP(sdpA,sdpB,spbC)(BBa_K1074009)operon is a killing factor of Bacillus subtilis.</p><br />
</div><br />
<div class="clear"></div><br />
<div class="part-col-2"><br />
<h1>PsdpRI+RBS+SDP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074011">BBa_K1074011</a></h2> <br />
<p>We construct this gene circuit as our kill switch to kill the engineered Bacillus subtilis for the safety purpose.Killing is mediated by the exported toxic protein SpbC. </p><br />
</div><br />
<div class="part-col-2"><br />
<br />
<br />
<br />
<h1>Pgrac promoter</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074012">BBa_K1074012</a></h2> <br />
<p>Prgac promoter(consisting of the groE promoter,the lacO operator and the gsiBSD sequence) allow induction by addition of ITPG. </p><br />
</div><br />
<br />
<br />
<br />
<div class="part-col-2"><br />
<h1>promoter sdpR/I</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074013">BBa_K1074013</a></h2> <br />
<p>This part is a sigma factor A-dependent promotor of the gene sdpR derived from the B.subtilis ,and it contains an operon for the signaling pathway of sdpc,a bacterial toxin of B.subtilis. </p><br />
</div><br />
<div class="part-col-2"><br />
<br />
<br />
<h1>SamyQ</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074014">BBa_K1074014</a></h2> <br />
<p>SamyQ is the signal peptide of the amyQ gene encoding an α-amylase in Bacillus subtilis WB800N. Fuse it with the recombinant proteins to obtain the secretion of recombinant proteins in Bacillus subtilis.</p><br />
</div><br />
<br />
<br />
<br />
<div class="part-col-2"><br />
<h1>RBS for Pgrac</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074015">BBa_K1074015</a></h2> <br />
<p>An efficient RBS for promoter Pgrac(BBa_K1074012) in Bacillus subtilis. We used it in all of our gene circuits containing promoter Pgrac. </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>RBS for Pctc</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074016">BBa_K1074016</a></h2> <br />
<p>This is an efficient RBS for promoter Pctc(BBa_K143010) in Bacillus subtilis. It's also an universal RBS for promoters in Bacillus subtilis. We also used it with the promoter PsdpRI(BBa_K1074013) in our project. </p><br />
</div><br />
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<div class="port-sidebar-border"><h>Project</h></div><br />
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<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails">Project Details</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails/Background">Background</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails/Design">Design</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Basic Experiment</a></div><br />
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</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/Notebook/TimelineTeam:USTC CHINA/Notebook/Timeline2013-09-27T14:11:15Z<p>Surine: </p>
<hr />
<div>{{USTC-China/hidden}}<br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
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<div id="title" align="center"><h>2013 USTC_CHINA iGEM Chronology</h></div><br />
<div class="main" align="left" style="background:#fff;border-radius:1em 1em 1em 1em;border:1px solid rgb(68,68,68);margin:20px auto 20px auto;"><br />
<div class="history"><br />
<div class="history-date"><br />
<ul><br />
<h2 class="first">2012</h2><br />
<br />
<li><br />
<h3><span>2012</span></h3><br />
<dl><br />
<dt><br />
</dt><br />
</dl><br />
</li><br />
<br />
<li class="green"><br />
<h3>Dec.15th<span>2012</span></h3><br />
<dl><br />
<dt>annual recruiting season<br />
<span>brought a large number of inquisitive mind <br />to USTC igem team. </span><br />
</dt><br />
</dl><br />
</li><br />
</ul><br />
</div><br />
<br />
<div class="history-date"><br />
<ul><br />
<h2 class="date02">2013</h2><br />
<li><br />
<h3>Jan.26th<span>2013</span></h3><br />
<dl><br />
<dt>systematic training begin<br />
<span>senior team members gave systematic training to the fresh<br /> and assigned responsibilities for every individual.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>Feb.17th<span>2013</span></h3><br />
<dl><br />
<dt>second training course<br />
<span>we held a simulated iGEM competition.<br /> Everyone was serious about the task he or she received,<br /> and gained a lot from the simulated competition.<br /> In the end, the team leader was elected by us.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>Mar.2nd<span>2013</span></h3><br />
<dl><br />
<dt>grouping and brain storming<br />
<span> All the members were divided into several groups<br /> according to each person's specialty and interest,<br /> and were motivated in the mobilization meeting.<br /> Everyone was ready for the coming activities.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>Mar.30th<span>2013</span></h3><br />
<dl><br />
<dt>Preliminary identified <br />several projects<br />
<span>algae produce H<sub>2</sub>, natural competence<br /> and magnetosome application <br />were preliminary identified as the promising projects.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>May.15th<span>2013</span></h3><br />
<dl><br />
<dt>SDI Conference <br />
<span>through heated discussion, we selected <br />optimization of blue-green algae produce H<sub>2</sub> as our subject.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>May.31th<span>2013</span></h3><br />
<dl><br />
<dt>halmatogenesis<br />
<span>A recently published paper has already done<br /> what we prepared to do, and we started to<br /> search another competitve project.</span><br />
</dt><br />
</dl><br />
</li><br />
<li class="green"><br />
<h3>June.5th<span>2013</span></h3><br />
<dl><br />
<dt>In situ transdermal vaccine<br />born<br />
</dt><br />
</dl><br />
</li><br />
<li class="green"><br />
<h3>July.10th-Apr.14th<span>2013</span></h3><br />
<dl><br />
<dt>experiment pet part<br />
<span>Introduce plasmid containing the GFP sequence into E.coli<br /><br />
Extract the plasmid after verified by PCR<br /><br />
Connect GFP gene with TD-1 via PCR<br /><br />
Connect the fragment with RBS and locus of restriction<br /> enzyme digestion via PCR<br /><br />
Digest the sequence and the plasmid <br />with same restriction endonuclease<br /><br />
Connect the sequence and the plasmid with DNA ligase<br /><br />
Verify the recombined plasmid by PCR<br /><br />
Sequence the plasmid<br /><br />
Introduce recombined plasmid into E.coli<br /><br />
Verify the bacterium by PCR<br /><br />
Induce protein expression<br /><br />
Verify the protein by SDS-page<br /><br />
Secret protein abundantly<br /><br />
Concentrate the protein via nickel column<br /><br />
Verify the protein by SDS-page<br /><br />
Transdermal experiments</span><br />
</dt><br />
</dl><br />
<br />
</li><br />
<li class="green"><br />
<h3>Apr.15th-Sept.10th<span>2013</span></h3><br />
<dl><br />
<dt>experiment B.subtilis part<br />
<span>Get the GFP sequence via PCR<br /><br />
Connect GFP gene with part of TD-1 via PCR<br /><br />
Connect the fragment with another part of TD-1 via PCR<br /><br />
Connect the fragment with promoter and <br />signal peptide via PCR<br /><br />
Digest the sequence and the plasmid with<br />same restriction endonuclease<br /><br />
Connect the sequence and the plasmid with DNA ligase<br /><br />
Verify the recombined plasmid by PCR<br /><br />
Sequence the plasmid<br /><br />
Introduce recombined plasmid into B.subtilis<br /><br />
Verify the bacterium by PCR<br /><br />
Induce protein expression<br /><br />
Concentrate the protein via TCA<br /><br />
Verify the protein by SDS-page<br /><br />
Secret protein abundantly<br /><br />
Concentrate by centrifuging<br /><br />
Verify the protein by SDS-page<br /><br />
Transdermal experiments</span><br />
</dt><br />
</dl><br />
</li><br />
<br />
<li class="green"><br />
<h3>Sept.14th<span>2013</span></h3><br />
<dl><br />
<dt>in vivo Transdermal antigen <br />antibody response validation<br />
</dt><br />
</dl><br />
</li><br />
</ul><br />
</div><br />
<br />
</div><br />
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</div><br />
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<br />
</body><br />
</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/Notebook/TimelineTeam:USTC CHINA/Notebook/Timeline2013-09-27T14:09:36Z<p>Surine: </p>
<hr />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
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<div id="title" align="center"><h>2013 USTC_CHINA iGEM Chronology</h></div><br />
<div class="main" align="left" style="background:#fff;border-radius:1em 1em 1em 1em;border:1px solid rgb(68,68,68);margin:20px auto 20px auto;"><br />
<div class="history"><br />
<div class="history-date"><br />
<ul><br />
<h2 class="first">2012</h2><br />
<br />
<li><br />
<h3><span>2012</span></h3><br />
<dl><br />
<dt><br />
</dt><br />
</dl><br />
</li><br />
<br />
<li class="green"><br />
<h3>Dec.15th<span>2012</span></h3><br />
<dl><br />
<dt>annual recruiting season<br />
<span>brought a large number of inquisitive mind <br />to USTC igem team. </span><br />
</dt><br />
</dl><br />
</li><br />
</ul><br />
</div><br />
<br />
<div class="history-date"><br />
<ul><br />
<h2 class="date02">2013</h2><br />
<li><br />
<h3>Jan.26th<span>2013</span></h3><br />
<dl><br />
<dt>systematic training begin<br />
<span>senior team members gave systematic training to the fresh<br /> and assigned responsibilities for every individual.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>Feb.17th<span>2013</span></h3><br />
<dl><br />
<dt>second training course<br />
<span>we held a simulated iGEM competition.<br /> Everyone was serious about the task he or she received,<br /> and gained a lot from the simulated competition.<br /> In the end, the team leader was elected by us.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>Mar.2nd<span>2013</span></h3><br />
<dl><br />
<dt>grouping and brain storming<br />
<span> All the members were divided into several groups<br /> according to each person's specialty and interest,<br /> and were motivated in the mobilization meeting.<br /> Everyone was ready for the coming activities.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>Mar.30th<span>2013</span></h3><br />
<dl><br />
<dt>Preliminary identified <br />several projects<br />
<span>algae produce H<sub>2</sub>, natural competence<br /> and magnetosome application <br />were preliminary identified as the promising projects.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>May.15th<span>2013</span></h3><br />
<dl><br />
<dt>SDI Conference <br />
<span>through heated discussion, we selected <br />optimization of blue-green algae produce H<sub>2</sub> as our subject.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>May.31th<span>2013</span></h3><br />
<dl><br />
<dt>halmatogenesis<br />
<span>A recently published paper had already done<br /> what we prepared to do, and we started to<br /> search another competitve project.</span><br />
</dt><br />
</dl><br />
</li><br />
<li class="green"><br />
<h3>June.5th<span>2013</span></h3><br />
<dl><br />
<dt>In situ transdermal vaccine<br />born<br />
</dt><br />
</dl><br />
</li><br />
<li class="green"><br />
<h3>July.10th-Apr.14th<span>2013</span></h3><br />
<dl><br />
<dt>experiment pet part<br />
<span>Introduce plasmid containing the GFP sequence into E.coli<br /><br />
Extract the plasmid after verified by PCR<br /><br />
Connect GFP gene with TD-1 via PCR<br /><br />
Connect the fragment with RBS and locus of restriction<br /> enzyme digestion via PCR<br /><br />
Digest the sequence and the plasmid <br />with same restriction endonuclease<br /><br />
Connect the sequence and the plasmid with DNA ligase<br /><br />
Verify the recombined plasmid by PCR<br /><br />
Sequence the plasmid<br /><br />
Introduce recombined plasmid into E.coli<br /><br />
Verify the bacterium by PCR<br /><br />
Induce protein expression<br /><br />
Verify the protein by SDS-page<br /><br />
Secret protein abundantly<br /><br />
Concentrate the protein via nickel column<br /><br />
Verify the protein by SDS-page<br /><br />
Transdermal experiments</span><br />
</dt><br />
</dl><br />
<br />
</li><br />
<li class="green"><br />
<h3>Apr.15th-Sept.10th<span>2013</span></h3><br />
<dl><br />
<dt>experiment B.subtilis part<br />
<span>Get the GFP sequence via PCR<br /><br />
Connect GFP gene with part of TD-1 via PCR<br /><br />
Connect the fragment with another part of TD-1 via PCR<br /><br />
Connect the fragment with promoter and <br />signal peptide via PCR<br /><br />
Digest the sequence and the plasmid with<br />same restriction endonuclease<br /><br />
Connect the sequence and the plasmid with DNA ligase<br /><br />
Verify the recombined plasmid by PCR<br /><br />
Sequence the plasmid<br /><br />
Introduce recombined plasmid into B.subtilis<br /><br />
Verify the bacterium by PCR<br /><br />
Induce protein expression<br /><br />
Concentrate the protein via TCA<br /><br />
Verify the protein by SDS-page<br /><br />
Secret protein abundantly<br /><br />
Concentrate by centrifuging<br /><br />
Verify the protein by SDS-page<br /><br />
Transdermal experiments</span><br />
</dt><br />
</dl><br />
</li><br />
<br />
<li class="green"><br />
<h3>Sept.14th<span>2013</span></h3><br />
<dl><br />
<dt>in vivo Transdermal antigen <br />antibody response validation<br />
</dt><br />
</dl><br />
</li><br />
</ul><br />
</div><br />
<br />
</div><br />
</div><br />
</div><br />
<br />
<br />
<div class="rightbar"><br />
<div class="port-sidebar-border"><h>Notebook</h></div><br />
<div class="clear"></div><br />
<div id="t1"><a class="active" href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">Timeline</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></div></div><br />
</div><br />
</div><br />
<br />
</body><br />
</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/Notebook/TimelineTeam:USTC CHINA/Notebook/Timeline2013-09-27T13:43:14Z<p>Surine: </p>
<hr />
<div>{{USTC-China/hidden}}<br />
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</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/">Modeling</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/DesignsofImmuneExperiments">Designs of Immune Experiments</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
<ul class="subs"><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Members</a></li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=USTC_CHINA">Profile</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Attributions">Attributions</a></li><br />
<li><a href="https://igem.org/2013_Judging_Form?id=1074#iGEM_Medals">Achievements</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Safety">Safety</a></li><br />
</ul><br />
<div id="tlogo"><img src="https://static.igem.org/mediawiki/2013/f/f8/2013ustc-china_T-VACCINE.png" width="100%" height="123" /><br />
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<div class="conbar2"><br />
<div class="leftbar"><br />
<div id="title" align="center"><h>2013 USTC_CHINA iGEM Chronology</h></div><br />
<div class="main" align="left" style="background:#fff;border-radius:1em 1em 1em 1em;border:1px solid rgb(68,68,68);margin:20px auto 20px auto;"><br />
<div class="history"><br />
<div class="history-date"><br />
<ul><br />
<h2 class="first">2012年</h2><br />
<br />
<li><br />
<h3><span>2012</span></h3><br />
<dl><br />
<dt><br />
</dt><br />
</dl><br />
</li><br />
<br />
<li class="green"><br />
<h3>Dec.15th<span>2012</span></h3><br />
<dl><br />
<dt>annual recruiting season<br />
<span>brought a large number of inquisitive mind <br />to USTC igem team. </span><br />
</dt><br />
</dl><br />
</li><br />
</ul><br />
</div><br />
<br />
<div class="history-date"><br />
<ul><br />
<h2 class="date02">2013年</h2><br />
<li><br />
<h3>Jan.26th<span>2013</span></h3><br />
<dl><br />
<dt>systematic training begin<br />
<span>senior team members gave systematic training to the fresh<br /> and assigned responsibilities for every individual.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>Feb.17th<span>2013</span></h3><br />
<dl><br />
<dt>second training course<br />
<span>we held a simulated iGEM competition.<br /> Everyone was serious about the task he or she received,<br /> and gained a lot from the simulated competition.<br /> In the end, the team leader was elected by us.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>Mar.2nd<span>2013</span></h3><br />
<dl><br />
<dt>grouping and brain storming<br />
<span> All the members were divided into several groups<br /> according to each person's specialty and interest,<br /> and were motivated in the mobilization meeting.<br /> Everyone was ready for the coming activities.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>Mar.30th<span>2013</span></h3><br />
<dl><br />
<dt>Preliminary identified <br />several projects<br />
<span>algae produce H2, natural competence<br /> and magnetosome application <br />were preliminary identified as the promising projects.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>May.15th<span>2013</span></h3><br />
<dl><br />
<dt>SDI Conference <br />
<span>through heated discussion, we selected <br />optimization of blue-green algae produce H2 as our subject.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>May.31th<span>2013</span></h3><br />
<dl><br />
<dt>halmatogenesis<br />
<span>A recently published paper has already done<br /> what we prepared to do, and we started to<br /> search another competitve project.</span><br />
</dt><br />
</dl><br />
</li><br />
<li class="green"><br />
<h3>June.5th<span>2013</span></h3><br />
<dl><br />
<dt>In situ transdermal vaccine<br />born<br />
</dt><br />
</dl><br />
</li><br />
<li class="green"><br />
<h3>July.10th-Apr.14th<span>2013</span></h3><br />
<dl><br />
<dt>experiment pet part<br />
<span>Introduce plasmid containing the GFP sequence into E.coli<br /><br />
Extract the plasmid after verified by PCR<br /><br />
Connect GFP gene with TD-1 via PCR<br /><br />
Connect the fragment with RBS and locus of restriction<br /> enzyme digestion via PCR<br /><br />
Digest the sequence and the plasmid <br />with same restriction endonuclease<br /><br />
Connect the sequence and the plasmid with DNA ligase<br /><br />
Verify the recombined plasmid by PCR<br /><br />
Sequence the plasmid<br /><br />
Introduce recombined plasmid into E.coli<br /><br />
Verify the bacterium by PCR<br /><br />
Induce protein expression<br /><br />
Verify the protein by SDS-page<br /><br />
Secret protein abundantly<br /><br />
Concentrate the protein via nickel column<br /><br />
Verify the protein by SDS-page<br /><br />
Transdermal experiments</span><br />
</dt><br />
</dl><br />
<br />
</li><br />
<li class="green"><br />
<h3>Apr.15th-Sept.10th<span>2013</span></h3><br />
<dl><br />
<dt>experiment B.subtilis part<br />
<span>Get the GFP sequence via PCR<br /><br />
Connect GFP gene with part of TD-1 via PCR<br /><br />
Connect the fragment with another part of TD-1 via PCR<br /><br />
Connect the fragment with promoter and <br />signal peptide via PCR<br /><br />
Digest the sequence and the plasmid with<br />same restriction endonuclease<br /><br />
Connect the sequence and the plasmid with DNA ligase<br /><br />
Verify the recombined plasmid by PCR<br /><br />
Sequence the plasmid<br /><br />
Introduce recombined plasmid into B.subtilis<br /><br />
Verify the bacterium by PCR<br /><br />
Induce protein expression<br /><br />
Concentrate the protein via TCA<br /><br />
Verify the protein by SDS-page<br /><br />
Secret protein abundantly<br /><br />
Concentrate by centrifuging<br /><br />
Verify the protein by SDS-page<br /><br />
Transdermal experiments</span><br />
</dt><br />
</dl><br />
</li><br />
<br />
<li class="green"><br />
<h3>Sept.14th<span>2013</span></h3><br />
<dl><br />
<dt>in vivo Transdermal antigen <br />antibody response validation<br />
</dt><br />
</dl><br />
</li><br />
</ul><br />
</div><br />
<br />
</div><br />
</div><br />
</div><br />
<br />
<br />
<div class="rightbar"><br />
<div class="port-sidebar-border"><h>Notebook</h></div><br />
<div class="clear"></div><br />
<div id="t1"><a class="active" href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">Timeline</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></div></div><br />
</div><br />
</div><br />
<br />
</body><br />
</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/Notebook/TimelineTeam:USTC CHINA/Notebook/Timeline2013-09-27T13:42:13Z<p>Surine: </p>
<hr />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
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<div id="title" align="center"><h>2013 USTC_CHINA iGEM Chronology</h></div><br />
<div class="main" align="left" style="background:#fff;border-radius:1em 1em 1em 1em;border:1px solid rgb(68,68,68);margin:20px auto 20px auto;"><br />
<div class="history"><br />
<div class="history-date"><br />
<ul><br />
<h2 class="first"><a href="#nogo">2012年</a></h2><br />
<br />
<li><br />
<h3><span>2012</span></h3><br />
<dl><br />
<dt><br />
</dt><br />
</dl><br />
</li><br />
<br />
<li class="green"><br />
<h3>Dec.15th<span>2012</span></h3><br />
<dl><br />
<dt>annual recruiting season<br />
<span>brought a large number of inquisitive mind <br />to USTC igem team. </span><br />
</dt><br />
</dl><br />
</li><br />
</ul><br />
</div><br />
<br />
<div class="history-date"><br />
<ul><br />
<h2 class="date02">2013年</h2><br />
<li><br />
<h3>Jan.26th<span>2013</span></h3><br />
<dl><br />
<dt>systematic training begin<br />
<span>senior team members gave systematic training to the fresh<br /> and assigned responsibilities for every individual.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>Feb.17th<span>2013</span></h3><br />
<dl><br />
<dt>second training course<br />
<span>we held a simulated iGEM competition.<br /> Everyone was serious about the task he or she received,<br /> and gained a lot from the simulated competition.<br /> In the end, the team leader was elected by us.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>Mar.2nd<span>2013</span></h3><br />
<dl><br />
<dt>grouping and brain storming<br />
<span> All the members were divided into several groups<br /> according to each person's specialty and interest,<br /> and were motivated in the mobilization meeting.<br /> Everyone was ready for the coming activities.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>Mar.30th<span>2013</span></h3><br />
<dl><br />
<dt>Preliminary identified <br />several projects<br />
<span>algae produce H2, natural competence<br /> and magnetosome application <br />were preliminary identified as the promising projects.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>May.15th<span>2013</span></h3><br />
<dl><br />
<dt>SDI Conference <br />
<span>through heated discussion, we selected <br />optimization of blue-green algae produce H2 as our subject.</span><br />
</dt><br />
</dl><br />
</li><br />
<li><br />
<h3>May.31th<span>2013</span></h3><br />
<dl><br />
<dt>halmatogenesis<br />
<span>A recently published paper has already done<br /> what we prepared to do, and we started to<br /> search another competitve project.</span><br />
</dt><br />
</dl><br />
</li><br />
<li class="green"><br />
<h3>June.5th<span>2013</span></h3><br />
<dl><br />
<dt>In situ transdermal vaccine<br />born<br />
</dt><br />
</dl><br />
</li><br />
<li class="green"><br />
<h3>July.10th-Apr.14th<span>2013</span></h3><br />
<dl><br />
<dt>experiment pet part<br />
<span>Introduce plasmid containing the GFP sequence into E.coli<br /><br />
Extract the plasmid after verified by PCR<br /><br />
Connect GFP gene with TD-1 via PCR<br /><br />
Connect the fragment with RBS and locus of restriction<br /> enzyme digestion via PCR<br /><br />
Digest the sequence and the plasmid <br />with same restriction endonuclease<br /><br />
Connect the sequence and the plasmid with DNA ligase<br /><br />
Verify the recombined plasmid by PCR<br /><br />
Sequence the plasmid<br /><br />
Introduce recombined plasmid into E.coli<br /><br />
Verify the bacterium by PCR<br /><br />
Induce protein expression<br /><br />
Verify the protein by SDS-page<br /><br />
Secret protein abundantly<br /><br />
Concentrate the protein via nickel column<br /><br />
Verify the protein by SDS-page<br /><br />
Transdermal experiments</span><br />
</dt><br />
</dl><br />
<br />
</li><br />
<li class="green"><br />
<h3>Apr.15th-Sept.10th<span>2013</span></h3><br />
<dl><br />
<dt>experiment B.subtilis part<br />
<span>Get the GFP sequence via PCR<br /><br />
Connect GFP gene with part of TD-1 via PCR<br /><br />
Connect the fragment with another part of TD-1 via PCR<br /><br />
Connect the fragment with promoter and <br />signal peptide via PCR<br /><br />
Digest the sequence and the plasmid with<br />same restriction endonuclease<br /><br />
Connect the sequence and the plasmid with DNA ligase<br /><br />
Verify the recombined plasmid by PCR<br /><br />
Sequence the plasmid<br /><br />
Introduce recombined plasmid into B.subtilis<br /><br />
Verify the bacterium by PCR<br /><br />
Induce protein expression<br /><br />
Concentrate the protein via TCA<br /><br />
Verify the protein by SDS-page<br /><br />
Secret protein abundantly<br /><br />
Concentrate by centrifuging<br /><br />
Verify the protein by SDS-page<br /><br />
Transdermal experiments</span><br />
</dt><br />
</dl><br />
</li><br />
<br />
<li class="green"><br />
<h3>Sept.14th<span>2013</span></h3><br />
<dl><br />
<dt>in vivo Transdermal antigen <br />antibody response validation<br />
</dt><br />
</dl><br />
</li><br />
</ul><br />
</div><br />
<br />
</div><br />
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<div class="port-sidebar-border"><h>Notebook</h></div><br />
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<div id="t1"><a class="active" href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Timeline">Timeline</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">Protocols</a></div></div><br />
</div><br />
</div><br />
<br />
</body><br />
</html></div>Surinehttp://2013.igem.org/Team:USTC_CHINA/PartsTeam:USTC CHINA/Parts2013-09-27T13:37:01Z<p>Surine: </p>
<hr />
<div>{{USTC-China/hidden}}<br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/KillSwitch">Kill Switch</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/Modeling/B.SubtilisCulture">B.Subtilis Culture</a></li><br />
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</ul><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice">Human Practice</a><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Communication" >Communication</a></li><br />
<li><a href="https://2013.igem.org/Team:USTC_CHINA/HumanPractice/Activity">Activity</a></li><br />
</ul><br />
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<li><a href="https://2013.igem.org/Team:USTC_CHINA/Team">Team</a><br />
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<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Project</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></div></div><br />
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<div class="part-col-1"><br />
<h1>TD1, Transdermal peptide</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074000">BBa_K1074000</a></h2><br />
<p>TD1 is a short synthetic peptide(ACSSSPSKHCG) identified by in vivo phage display, facilitated efficient transdermal protein delivery through intact skin. Studies suggested that the peptide creates a transient opening in the skin barrier to enable macromolecular material to reach systemic circulation.</p><br />
</div><br />
<br />
<div class="part-col-1"><br />
<h1>Pgrac+RBS+SamyQ+TD1<br>+GFP</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074006">BBa_K1074006</a></h2><br />
<p>This is the main circuit of our project to allow high expression of target protein(antigen,adjuvant).GFP can be substituted by various proteins via a modular PCR or standard cut/ligation method.</p><br />
</div><br />
<div class="clear"></div><br />
<div class="part-col-2"><br />
<h1>PHT43</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074001">BBa_K1074001</a></h2><br />
<p>PHT43 is a E.coli-B.subtilis shuttle vector allowing high-level expression of secreted proteins in B.subtilis. </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>HBsAg</h1><br />
<h2><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1074002">BBa_K1074002</a></h2> <br />
<p>HBsAg is the surface antigen of the hepatitis B virus (HBV). It indicates current hepatitis B infection.</p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>Overview</h1><br />
<p>In our world, billions of people suffer from contagion, however, only part of them can be prevented by proper vaccine. Many disadvantages limit the usage of traditional vaccine especially in developing countries. </p><br />
</div><br />
<br />
<div class="part-col-2"><br />
<h1>Overview</h1><br />
<p>In our world, billions of people suffer from contagion, however, only part of them can be prevented by proper vaccine. Many disadvantages limit the usage of traditional vaccine especially in developing countries. </p><br />
</div><br />
<div class="clear"></div><br />
<div class="part-col-2"><br />
<h1>Overview</h1><br />
<p>In our world, billions of people suffer from contagion, however, only part of them can be prevented by proper vaccine. Many disadvantages limit the usage of traditional vaccine especially in developing countries. </p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Overview</h1><br />
<p>In our world, billions of people suffer from contagion, however, only part of them can be prevented by proper vaccine. Many disadvantages limit the usage of traditional vaccine especially in developing countries. </p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Overview</h1><br />
<p>In our world, billions of people suffer from contagion, however, only part of them can be prevented by proper vaccine. Many disadvantages limit the usage of traditional vaccine especially in developing countries. </p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Overview</h1><br />
<p>In our world, billions of people suffer from contagion, however, only part of them can be prevented by proper vaccine. Many disadvantages limit the usage of traditional vaccine especially in developing countries. </p><br />
</div><br />
<div class="clear"></div><br />
<div class="part-col-2"><br />
<h1>Overview</h1><br />
<p>In our world, billions of people suffer from contagion, however, only part of them can be prevented by proper vaccine. Many disadvantages limit the usage of traditional vaccine especially in developing countries. </p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Overview</h1><br />
<p>In our world, billions of people suffer from contagion, however, only part of them can be prevented by proper vaccine. Many disadvantages limit the usage of traditional vaccine especially in developing countries. </p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Overview</h1><br />
<p>In our world, billions of people suffer from contagion, however, only part of them can be prevented by proper vaccine. Many disadvantages limit the usage of traditional vaccine especially in developing countries. </p><br />
</div><br />
<div class="part-col-2"><br />
<h1>Overview</h1><br />
<p>In our world, billions of people suffer from contagion, however, only part of them can be prevented by proper vaccine. Many disadvantages limit the usage of traditional vaccine especially in developing countries. </p><br />
</div><br />
<div class="clear"></div><br />
</div><br />
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<div class="port-sidebar-border"><h>Project</h></div><br />
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<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Overview">Overview</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails">Project Details</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails/Background">Background</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/ProjectDetails/Design">Design</a></div><br />
<div id="t1"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Results</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results">Basic Experiment</a></div><br />
<div id="t2"><a href="https://2013.igem.org/Team:USTC_CHINA/Project/Results/FurthurWork">Furthur Work</a></div><br />
<div id="t1"><a class="active" href="https://2013.igem.org/Team:USTC_CHINA/Parts">Parts</a></div><br />
</div></div></div> <br />
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</html></div>Surine