The band marked in red disappeared is pSB1C3-gfp-luxI/BL21.The LuxR, GFP and AiiA are the same of size (marked in yellow). However the band of AiiA was not clear. Many target proteins were misfolded. Therefore the culture condition should be optimized.
The band marked in red disappeared is pSB1C3-gfp-luxI/BL21.The LuxR, GFP and AiiA are the same of size (marked in yellow). However the band of AiiA was not clear. Many target proteins were misfolded. Therefore the culture condition should be optimized.
We had transformed triple plasmids into one E.coli and the DNA gel picture indicated it was ligated successfully. So now we were very eager to know if our circuits could work well in E.coli.
We had transformed triple plasmids into one E.coli and the DNA gel picture indicated it was ligated successfully. So now we were very eager to know if our circuits could work well in E.coli.
Furthermore, we wanted to know that if our target proteins expressed by our constructed gene circuits, in other words, pSB1C3-gfp-luxI and pSB13T5-aiiA were aggregated in precipitation or supernatant. So we run SDS-PAGE using BL21(blank control), three cells transformed into different single plasmids(aiiA, ndh and gfp), one cell transformed into double plasmids(aiiA and gfp) and our E.coli transformed into triple plasmids in three forms respectively, whole cell, precipitation and supernatant.
Furthermore, we wanted to know that if our target proteins expressed by our constructed gene circuits, in other words, pSB1C3-gfp-luxI and pSB13T5-aiiA were aggregated in precipitation or supernatant. So we run SDS-PAGE using BL21(blank control), three cells transformed into different single plasmids(aiiA, ndh and gfp), one cell transformed into double plasmids(aiiA and gfp) and our E.coli transformed into triple plasmids in three forms respectively, whole cell, precipitation and supernatant.
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Line 182:
Data analysis:
Data analysis:
The band marked in red was not the target band. Maybe it is the complex of AHL-LuxI. The target proteins of pSB1C3-gfp-luxI have been expressed while the target proteins of pSB3T5-aiiA and pSB4K5-ndh did not be expressed because the promoter should be induced by AHL.
The band marked in red was not the target band. Maybe it is the complex of AHL-LuxI. The target proteins of pSB1C3-gfp-luxI have been expressed while the target proteins of pSB3T5-aiiA and pSB4K5-ndh did not be expressed because the promoter should be induced by AHL.
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</textarea> <br> </td> </tr> </tbody></table> </td> </tr> </tbody></table> <div class="M1_exp_stepInfo" title="5. Confirmation of the expression level cultured at 30℃(8.31)" id="sb7s7" > <button class="M1_exp_step_name_button" ><input value="5. Confirmation of the expression level cultured at 30℃(8.31)" class="M1_exp_step_name"></button> <span class="M1_exp_step_time">2013/9/26 22:09:10--Tang Chun</span> </div> <table class="M1_exp_stepBoard"> <tbody><tr class="M1_exp_stepItem"> <td class="M1_exp_item_time" id="ib7s7i0">2013/9/26 22:09:28</td> <td class="M1_exp_item_content"> <table id="i7s7i0" class="C1"> <tbody><tr> <td class="C1_td"> <img id="i7s7i00" width="500px" src="http://trysomething-block.stor.sinaapp.com/t14i9s7i05.png"><br> <form id="i7s7i04" target="M7" action="img.php" method="post" enctype="multipart/form-data"></form> <input name="img" type="file" id="i7s7i02" form="i7s7i04" class="C1_button1" required="required" disabled="" style="background-color: white;"> <input name="pname" type="hidden" value="t14" form="i7s7i04"> <input name="id" type="hidden" value="i7s7i0" form="i7s7i04"> <input name="oldgraph" type="hidden" id="i7s7i05" form="i7s7i04" value="t14i9s7i05.png"> <input id="i7s7i06" type="submit" value="submit" form="i7s7i04" class="C1_button2" disabled="" style="background-color: white;"> <div id="i7s7i03" class="M1_exp_img"></div> </td> <td class="C1_td_text"> <textarea id="i7s7i01" class="C1_text" disabled="" style="background-color: white;"> We were suggested that DH5α has a better viability and a higher protein expression. So we maybe there's some mistakes in our strains which cause our couldn't get a expected microfluidic experiment result. So we compared two identical groups cultured at different degrees, 30℃ and 37℃.
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</textarea> <br> </td> </tr> </tbody></table> </td> </tr> </tbody></table> <div class="M1_exp_stepInfo" title="5. Confirmation of the expression level cultured at 30℃(8.31)" id="sb7s7" > <button class="M1_exp_step_name_button" ><input value="5. Confirmation of the expression level cultured at 30℃(8.31)" class="M1_exp_step_name"></button> <span class="M1_exp_step_time">2013/9/26 22:09:10--Tang Chun</span> </div> <table class="M1_exp_stepBoard"> <tbody><tr class="M1_exp_stepItem"> <td class="M1_exp_item_time" id="ib7s7i0">2013/9/26 22:09:28</td> <td class="M1_exp_item_content"> <table id="i7s7i0" class="C1"> <tbody><tr> <td class="C1_td"> <img id="i7s7i00" width="500px" src="http://trysomething-block.stor.sinaapp.com/t14i9s7i05.png"><br> <form id="i7s7i04" target="M7" action="img.php" method="post" enctype="multipart/form-data"></form> <input name="img" type="file" id="i7s7i02" form="i7s7i04" class="C1_button1" required="required" disabled="" style="background-color: white;"> <input name="pname" type="hidden" value="t14" form="i7s7i04"> <input name="id" type="hidden" value="i7s7i0" form="i7s7i04"> <input name="oldgraph" type="hidden" id="i7s7i05" form="i7s7i04" value="t14i9s7i05.png"> <input id="i7s7i06" type="submit" value="submit" form="i7s7i04" class="C1_button2" disabled="" style="background-color: white;"> <div id="i7s7i03" class="M1_exp_img"></div> </td> <td class="C1_td_text"> <textarea id="i7s7i01" class="C1_text" disabled="" style="background-color: white;"> We were suggested that DH5α has a better viability and a higher protein expression. So there's some mistakes in our strains which cause us couldn't get a expected microfluidic experiment result. So we compared two identical groups cultured at different degrees, 30℃ and 37℃.
Data analysis:
Data analysis:
E.coli maybe cultured at 30℃ had a stronger expression of protein from picture 1 but we were confused by picture 2 cause it indicated that expression at 30℃ is not larger than expression at 37℃.</textarea> <br> </td> </tr> </tbody></table> </td> </tr> <tr class="M1_exp_stepItem"> <td class="M1_exp_item_time" id="ib7s7i1">2013/9/26 22:09:40</td> <td class="M1_exp_item_content"> <table id="i7s7i1" class="C1"> <tbody><tr> <td class="C1_td"> <img id="i7s7i10" src="http://trysomething-block.stor.sinaapp.com/t14i9s7i16.png"><br> <form id="i7s7i14" target="M7" action="img.php" method="post" enctype="multipart/form-data"></form> <input name="img" type="file" id="i7s7i12" form="i7s7i14" class="C1_button1" required="required" disabled="" style="background-color: white;"> <input name="pname" type="hidden" value="t14" form="i7s7i14"> <input name="id" type="hidden" value="i7s7i1" form="i7s7i14"> <input name="oldgraph" type="hidden" id="i7s7i15" form="i7s7i14" value="t14i9s7i16.png"> <input id="i7s7i16" type="submit" value="submit" form="i7s7i14" class="C1_button2" disabled="" style="background-color: white;"> <div id="i7s7i13" class="M1_exp_img"></div> </td> <td class="C1_td_text"> <textarea id="i7s7i11" class="C1_text" disabled="" style="background-color: white;">Future plan:
E.coli maybe cultured at 30℃ had a stronger expression of protein from picture 1 but we were confused by picture 2 cause it indicated that expression at 30℃ is not larger than expression at 37℃.</textarea> <br> </td> </tr> </tbody></table> </td> </tr> <tr class="M1_exp_stepItem"> <td class="M1_exp_item_time" id="ib7s7i1">2013/9/26 22:09:40</td> <td class="M1_exp_item_content"> <table id="i7s7i1" class="C1"> <tbody><tr> <td class="C1_td"> <img id="i7s7i10" src="http://trysomething-block.stor.sinaapp.com/t14i9s7i16.png"><br> <form id="i7s7i14" target="M7" action="img.php" method="post" enctype="multipart/form-data"></form> <input name="img" type="file" id="i7s7i12" form="i7s7i14" class="C1_button1" required="required" disabled="" style="background-color: white;"> <input name="pname" type="hidden" value="t14" form="i7s7i14"> <input name="id" type="hidden" value="i7s7i1" form="i7s7i14"> <input name="oldgraph" type="hidden" id="i7s7i15" form="i7s7i14" value="t14i9s7i16.png"> <input id="i7s7i16" type="submit" value="submit" form="i7s7i14" class="C1_button2" disabled="" style="background-color: white;"> <div id="i7s7i13" class="M1_exp_img"></div> </td> <td class="C1_td_text"> <textarea id="i7s7i11" class="C1_text" disabled="" style="background-color: white;">Future plan:
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Line 213:
There is a difference between N and N+A：
There is a difference between N and N+A：
The band marked in green/red shows that E.coli cultured has a stronger expression of protein when added AHL?
The band marked in green/red shows that E.coli cultured has a stronger expression of protein when added AHL?
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</textarea> <br> </td> </tr> </tbody></table> </td> </tr> </tbody></table> <div class="M1_exp_stepInfo" title="10.Characterization of several parts using additional AHL .(9.27)" id="sb7s12"> <button class="M1_exp_step_name_button"><input value="10.Characterization of several parts using additional AHL .(9.27)" class="M1_exp_step_name"></button> <span class="M1_exp_step_time">2013/9/28 08:44:02--Chun Tang</span> </div> <table class="M1_exp_stepBoard"> <tbody><tr class="M1_exp_stepItem"> <td class="M1_exp_item_time" id="ib7s12i0">2013/9/28 08:44:22</td> <td class="M1_exp_item_content"> <table id="i7s12i0" class="C1"> <tbody><tr> <td class="C1_td"> <img id="i7s12i00" src="http://trysomething-block.stor.sinaapp.com/t14i7s12i0sds1.png"><br> <form id="i7s12i04" target="M7" action="img.php" method="post" enctype="multipart/form-data"></form> <input name="img" type="file" id="i7s12i02" form="i7s12i04" class="C1_button1" required="required" disabled=""> <input name="pname" type="hidden" value="t14" form="i7s12i04"> <input name="id" type="hidden" value="i7s12i0" form="i7s12i04"> <input name="oldgraph" type="hidden" id="i7s12i05" form="i7s12i04" value="t14i7s12i0sds1.png"> <input id="i7s12i06" type="submit" value="submit" form="i7s12i04" class="C1_button2" disabled=""> <div id="i7s12i03" class="M1_exp_img"></div> </td> <td class="C1_td_text"> <textarea id="i7s12i01" class="C1_text"> We continued to characterize our biobricks using additional AHL. These plasmids included sfgfp, sfgfp aiiA, sfgfp aiiA ndh. SDS-PAGE gel pictures are shown as follows.
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</textarea> <br> </td> </tr> </tbody></table> </td> </tr> </tbody></table> <div class="M1_exp_stepInfo" title="10.Characterization of several parts using additional AHL .(9.27)" id="sb7s12"> <button class="M1_exp_step_name_button"><input value="10.Characterization of several parts using additional AHL .(9.27)" class="M1_exp_step_name"></button> <span class="M1_exp_step_time">2013/9/28 08:44:02--Chun Tang</span> </div> <table class="M1_exp_stepBoard"> <tbody><tr class="M1_exp_stepItem"> <td class="M1_exp_item_time" id="ib7s12i0">2013/9/28 08:44:22</td> <td class="M1_exp_item_content"> <table id="i7s12i0" class="C1"> <tbody><tr> <td class="C1_td"> <img id="i7s12i00" src="http://trysomething-block.stor.sinaapp.com/t14i7s12i0sds1.png"><br> <form id="i7s12i04" target="M7" action="img.php" method="post" enctype="multipart/form-data"></form> <input name="img" type="file" id="i7s12i02" form="i7s12i04" class="C1_button1" required="required" disabled=""> <input name="pname" type="hidden" value="t14" form="i7s12i04"> <input name="id" type="hidden" value="i7s12i0" form="i7s12i04"> <input name="oldgraph" type="hidden" id="i7s12i05" form="i7s12i04" value="t14i7s12i0sds1.png"> <input id="i7s12i06" type="submit" value="submit" form="i7s12i04" class="C1_button2" disabled=""> <div id="i7s12i03" class="M1_exp_img"></div> </td> <td class="C1_td_text"> <textarea id="i7s12i01" class="C1_text"> We continued to characterize our biobricks using additional AHL. These plasmids included sfgfp, sfgfp-aiiA, sfgfp-aiiA-ndh. SDS-PAGE gel pictures are shown as follows.
"
