http://2013.igem.org/wiki/index.php?title=Special:Contributions/Sabrijamal&feed=atom&limit=50&target=Sabrijamal&year=&month=2013.igem.org - User contributions [en]2024-03-28T22:33:58ZFrom 2013.igem.orgMediaWiki 1.16.5http://2013.igem.org/Team:Uppsala/advisorsTeam:Uppsala/advisors2013-10-28T10:54:32Z<p>Sabrijamal: </p>
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<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
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<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">p-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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</ul></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">Kinetic model</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/toxicity-model">Toxicity model</a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/advisors">Advisors</a></li><br />
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</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/bioart">BioArt</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/LactonutritiousWorld">A LactoWorld</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/killswitches">Killswitches</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/realization">Patent</a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a></li> <br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/protocols">Protocols</a></li><br />
</ul></li><br />
</ul><br />
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<h1>Supervisors</h1><br />
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<h1>Anders Virtanen</h1><br />
<p>Anders Virtanen is a professor at the Department of Cell and Molecular Biology and the director of the Uppsala RNA Research Centre (URRC). Anders studies primarily enzymes involved in regulating fate and function of the plethora of RNA molecules in the cell. Another goal is to develop novel antibiotics targeting RNA molecules.</p><br />
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<h1>Anthony Forster</h1><br />
<p>Anthony Forster is a professor at the Department of Cell and Molecular Biology and a member of the Uppsala RNA Research Centre (URRC). He is researching synthetic biology, protein synthesis and drug discovery, and is currently working towards the synthesis of a minimal cell.</p><br />
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<h1>Mikael Nissbeck</h1><br />
<p>PhD student at Uppsala RNA Research Center (URRC), Department of Cell and Molecular biology, at Uppsala University since 2012 and is currently working on the human enzyme Poly(A)-specific ribonuclease (PARN) in the Anders Virtanen group at the Uppsala Biomedical Center.</p><br />
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<h1> Abhishek Srivastava </h1><br />
<p>Postdoc at Chemical Biology Group, Department of Cell<br />
and Molecular biology, at Uppsala University since 2012 and is<br />
currently working on ribonuclease P in Leif Kirsebom group at the <br />
Uppsala Biomedical Center.</p><br />
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<img class="pic" src="https://static.igem.org/mediawiki/2013/1/17/Uppsala2013_roos.jpg"><br />
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<h1>Stefan Roos</h1><br />
<p>Stefan Roos is an associate professor at the Department of Microbiology at Swedish University of Agricultural Sciences in Uppsala. Stefan works with genomics, physiology and ecology of probiotic lactobacilli. The primary goals are to understand how those bacteria interact with their host and to design probiotic products with improved activity.</p><br />
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<h1> Sethu Madhava Rao Gunja </h1><br />
<p>Research Assistant at Chemical Biology group, Department of Cell and Molecular Biology in Uppsala University since 2012 and is currently working on Physiological characterisation of human enzyme Poly(A)-specific ribonuclease (PARN) in the Anders Virtanen group at the Uppsala Biomedical Center.</p><br />
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<h1>Erik Gullberg</h1><br />
<p>PhD student in Medical Bacteriology at the Department of Medical Biochemistry and Microbiology at Uppsala University since 2009 and is currently working on antibiotic resistance in bacteria in the Dan Andersson group at the Uppsala Biomedical Center. So far he has published articles in PLoS Pathogens and Molecular Microbiology.</p><br />
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<img class="pic" src="https://static.igem.org/mediawiki/2013/7/7c/Uppsala2013_lundin.jpg"><br />
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<h1>Erik Lundin</h1><br />
<p>PhD student in Medical Bacteriology at the Department of Medical Biochemistry and Microbiology at Uppsala University since 2013. Erik Lundin is currently working on bacterial evolution where he studies evolution of new genes and functions. Erik works in Dan Andersson group at the Uppsala Biomedical Center. </p><br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/teamTeam:Uppsala/team2013-10-04T21:22:08Z<p>Sabrijamal: </p>
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<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/advisors">Advisors</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a></li> <br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/protocols">Protocols</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
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<h1 class="main-title"> Team iGEM Uppsala 2013 </h1><br />
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Team Uppsala iGEM is competing for the fifth consecutive year. This time we are a group of 26 under and overgraduate students in the fields of biotechnology and microbiology. We study at Uppsala University and it’s academic sister Swedish University of Agricultural Sciences. Both are located in Uppsala, once the capital of Sweden and center of norse religion; and today home to the oldest university in scandinavia and a big life science cluster. Our lab is located at Uppsala Biomedical Center.<br />
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Because we are a large team we have decided to divide ourselves into three groups. Two groups have focused on metabolic engineering and one group focused on creating a new probiotic chassi. <br />
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<img class="team-pic-template" src="https://static.igem.org/mediawiki/2013/f/f2/Team_pic_uppsala4.png"><br />
<i class="team-pic-text">Team iGEM Uppsala enjoying a swedish summer sunset outside Uppsala Biomedical Center. </i><br />
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<b>Friday Night Practice Sessions</b><br />
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Please use this sign-up sheet to book a 30-minute time slot to practice your presentation. Pick up your favorite bacterium genus and appropriate time slot, log in with your user account and edit this wiki page. <br />
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Ten practice rooms will be available from 5pm to 8pm on Friday with 30-minute time slots. Every team should be able to fit in, but if you really don't have a choice and need to practice your presentation after 8pm or during a time slot that's already completed, we'll try to come up with something. Please <a href ="https://2013.igem.org/Europe/Contact">contact us</a> in case of need.<br />
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Every practice room will be equipped with a standard video projector. Please make sure to bring your own computer and appropriate cables to set things up.<br />
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And last but not least, we are pretty sure that you would not enjoy losing time when you get to the practice room and find out that you have to wait because the previous team is late on schedule, so be sure arrive and leave on time. Thank you.<br />
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{| class="wikitable"<br />
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! |<br />
! colspan="10" style="font-style:normal;"| Practice Rooms <br />
|- style="height:25px;"<br />
! | !! | Agrobacterium !! Bacillus !! | Collimonas !! | Dickeya !! | Escherichia !! | Fusobacter !! | Gemminibacter !! | Helicobacter !! | Ignicoccus !! | Jannaschia<br />
|-<br />
! scope="row"| 5:00-5:30pm<br />
| Braunschweig || Newcastle || SDU-Denmark || TU-Munich || METU || Bielefeld-Germany || G1 || Leeds || DTU-Denmark || INSA Toulouse<br />
|-<br />
! scope="row"| 5:30-6:00pm<br />
| A2 || Göttingen || C2 || KU_Leuven || UGent || F2 || Paris_Saclay || UniSalento Lecce || I2 || J2<br />
|-<br />
! scope="row"| 6:00-6:30pm<br />
| Uppsala || B3 || Warsaw || Valencia_Biocampus || Exeter || TU Eindhoven || G3 || NRP-UEA-Norwich || Imperial College London || J3<br />
|-<br />
! scope="row"| 6:30-7:00pm<br />
| A4 || B4 || Kent || D4 || BGU_Israel || ATOMS-Turkiye || G4 || H4 || I4 || UCL post-grad<br />
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! scope="row"| 7:00-7:30pm<br />
| A5 || Bonn || C5 || Grenoble-EMSE-LSU || ETH Zurich || Wageningen UR || G5 || Evry || EPF_Lausanne || Freiburg<br />
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! scope="row"| 7:30-8:00pm<br />
| UNIK_Copenhagen || UNITN-Trento || Poznan-BioInf || Paris_Bettencourt || Manchester || Groningen || Leicester || TU-Delft || Dundee || UCL undergrad<br />
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|}</div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/yoghurtTeam:Uppsala/yoghurt2013-10-04T20:34:07Z<p>Sabrijamal: </p>
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
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<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/advisors">Advisors</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a></li> <br />
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<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/protocols">Protocols</a></li><br />
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<h1 class="subheader">The next generation of synthetic probiotics</h1><br />
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<p> iGEM Uppsala proudly presents a new way of creating nutritious food, by producing healthy substances in probiotic bacteria. As a part of our human practice project we have made a concept yoghurt product. Of course, as a medium for carrying our bacteria we are not only restricted to yoghurt. These probiotics can be provided in a juice, a pill and many other different foods.<br><br />
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The reasons for doing this concept product are as following:</p><br />
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<li class="yoghurt-text-list">We felt that we early on in the project wanted to have a clear vision on what our final goal would be.</li><br />
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<li class="yoghurt-text-list">This concept product makes it easier for us to explain for the general public what we are doing. We have seen that media and people are more interested in our project when they can envision exactly what we are trying to accomplish.</li><br />
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<li class="yoghurt-text-list">By creating this yoghurt, we can use this as a tool to start a debate and discuss future applications of synthetic biology.</li><br><br />
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<p>Our goal with this yoghurt is to make probiotic cultures produce all of our daily needs. When we use probiotics to produce nutritions, we do exactly what nature has done for us throughout all ages. With this, we can spare our nature, by going into the future with a new way of producing our food.<br />
This is a yoghurt that contains everything in one drink. With this, people can forget about expensive pills and supplements.<br><br><br />
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Our nutritions are for example produced in the healthy probiotic culture lactobacillus reuteri, which in themselves, are very good for our wellbeing.<br><br><br />
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In the end, all of the genes producing these different nutritions will be incorporated in the chromosome of our probiotics. This will eliminate the need for antibiotic resistance markers on our final product, thus eliminating the risk of spreading antibiotic resistance in our nature and ingesting them.</p> <br />
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<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
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<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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</ul></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/advisors">Advisors</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
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</ul></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/protocols">Protocols</a></li><br />
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<h1 class="subheader">The next generation of synthetic probiotics</h1><br />
<br />
<p> iGEM Uppsala proudly presents a new way of creating nutritious food – by producing healthy substances in probiotic bacteria. As a part of our human practice project we have made a concept yoghurt product. Of course, as a medium for carrying our bacteria we are not only restricted to yoghurt. These probiotics can be provided in a juice, a pill and many other different foods.<br><br />
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The reasons for doing this concept product are as following:</p><br />
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<li class="yoghurt-text-list">We felt that we early on in the project wanted to have a clear vision on what our final goal would be.</li><br />
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<li class="yoghurt-text-list">This concept product makes it easier for us to explain for the general public what we are doing. We have seen that media and people are more interested in our project when they can envision exactly what we are trying to accomplish.</li><br />
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<li class="yoghurt-text-list">By creating this yoghurt, we can use this as a tool to start a debate and discuss future applications of synthetic biology.</li><br><br />
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<br> <br />
<p>Our goal with this yoghurt is to make probiotic cultures produce all of our daily needs. When we use probiotics to produce nutritions, we do exactly what nature has done for us throughout all ages. With this, we can spare our nature, by going into the future with a new way of producing our food.<br />
This is a yoghurt that contains everything in one drink. With this, people can forget about expensive pills and supplements.<br><br><br />
<br />
Our nutritions are for example produced in the healthy probiotic culture lactobacillus reuteri, which in themselves, are very good for our wellbeing.<br><br><br />
<br />
In the end, all of the genes producing these different nutritions will be incorporated in the chromosome of our probiotics. This will eliminate the need for antibiotic resistance markers on our final product, thus eliminating the risk of spreading antibiotic resistance in our nature and ingesting them.</p> <br />
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<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
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<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/advisors">Advisors</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/protocols">Protocols</a></li><br />
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<h1 class="main-title">Collaboration</h1><br />
<br />
All of the work that was done during this project was both planned and executed by our iGEM team members. It all started of with some brainstorming sessions that began in the middle of May, although the actual lab work began in the beginning of June. <br />
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<br><br />
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During the summer we have worked very closely with the <a href="https://2013.igem.org/Team:Linkoping_Sweden"> Linköpings iGEM team 2013 </a>. Since it was their first year to participate in the competition we decided to do our best to provide them with any assistance they needed, such as providing them with parts, knowledge etc.<br />
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In addition to assisting Linköpings iGEM team we have also sent biobricks to the following iGEM teams:<br />
<br><br />
<br><br />
<br />
iGEM 2013 <a href="https://2013.igem.org/Team:UT-Tokyo"> UT-Tokyo: </a> <br><br />
<a href="http://parts.igem.org/Part:BBa_K592010"> BBa_K592010 </a> - amilGFP, yellow chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K864401"> BBa_K864401 </a> - aeBlue, blue chromoprotein <br><br />
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<br><br />
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iGEM 2013 <a href="https://2013.igem.org/Team:Braunschweig"> Braunschweig: </a> <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033930">BBa_K1033930</a> - amilCP, blue/purple chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K592010"> BBa_K592010 </a> - amilGFP, yellow chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592012"> BBa_K592012 </a> - eforRed, red/pink chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592011"> BBa_K592011 </a> - cjBlue, green chromoprotein<br><br />
<a href="http://parts.igem.org/Part:BBa_K864401"> BBa_K864401 </a> - aeBlue, blue chromoprotein <br><br />
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<br><br />
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iGEM 2013 <a href="https://2013.igem.org/Team:Tsinghua"> Tsinghua: </a> <br><br />
<a href="http://parts.igem.org/Part:BBa_K592010"> BBa_K592010 </a> - amilGFP, yellow chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592012"> BBa_K592012 </a> - eforRed, red/pink chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592011"> BBa_K592011 </a> - cjBlue, green chromoprotein<br><br />
<a href="http://parts.igem.org/Part:BBa_K864401"> BBa_K864401 </a> - aeBlue, blue chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033930">BBa_K1033930</a> - amilCP, blue/purple chromoprotein (incl RBS) <br><br />
<br />
<br><br />
<br />
iGEM 2013 <a href="https://2013.igem.org/Team:CU-Boulder"> CU Boulder: </a> <br><br />
<a href="http://parts.igem.org/Part:BBa_K592010"> BBa_K592010 </a> - amilGFP, yellow chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592012"> BBa_K592012 </a> - eforRed, red/pink chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592011"> BBa_K592011 </a> - cjBlue, green chromoprotein<br><br />
<a href="http://parts.igem.org/Part:BBa_K864401"> BBa_K864401 </a> - aeBlue, blue chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033930">BBa_K1033930</a> - amilCP, blue/purple chromoprotein (incl RBS) <br><br />
<br />
<br><br />
<br />
iGEM 2013 <a href="https://2013.igem.org/Team:uOttawa"> uOttawa: </a> <br><br />
<a href="http://parts.igem.org/Part:BBa_K592010"> BBa_K592010 </a> - amilGFP, yellow chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592012"> BBa_K592012 </a> - eforRed, red/pink chromoprotein <br><br />
<br />
<br><br />
<br />
iGEM 2013 <a href="https://2013.igem.org/Team:HokkaidoU_Japan"> HokkaidoU: </a> <br><br />
<a href="http://parts.igem.org/Part:BBa_K592009"> BBa_K592009 </a> - amilCP, blue chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033931">BBa_K1033931</a> - amilGFP, yellow chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K592010"> BBa_K592010 </a> - amilGFP, yellow chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592012"> BBa_K592012 </a> - eforRed, red/pink chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592011"> BBa_K592011 </a> - cjBlue, green chromoprotein<br><br />
<a href="http://parts.igem.org/Part:BBa_K864401"> BBa_K864401 </a> - aeBlue, blue chromoprotein <br><br />
<br />
<br><br />
<br />
iGEM 2013 <a href="https://2013.igem.org/Team:TU_Darmstadt"> TU Darmstadt: </a> <br><br />
<a href="http://parts.igem.org/Part:BBa_K592012"> BBa_K592012 </a> - eforRed, red/pink chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592011"> BBa_K592011 </a> - cjBlue, green chromoprotein<br><br />
<a href="http://parts.igem.org/Part:BBa_K1033929">BBa_K1033929</a> - aeBlue, blue chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033928">BBa_K1033928</a> - aeBlue, blue chromoprotein (incl RBS, J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033903"> BBa_K1033903 </a> - tsPurple, purple chromoprotein (incl RBS, J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033930">BBa_K1033930</a> - amilCP, blue/purple chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033931">BBa_K1033931</a> - amilGFP, yellow chromoprotein (incl RBS) <br><br />
<br />
<br><br />
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<h1> A special thanks </h1><br />
<br />
We would also like to thank other sources that have helped us, such as the <a href="https://2010.igem.org/Team:Slovenia"> Slovenian iGEM team 2010 </a> that suppied us with the genes needed for the production of carotenoid metabolites, <a href="https://2012.igem.org/Team:TU_Munich"> TU-Munichs iGEM team 2012 </a> that provided us with the gene 4CL for the production of 4-coumaryl-CoA, last but not least we would also like to thank the School of chemical and Biomolecular Engineering, Cornell University for providing us with the genes involved in the production of the phenylpropanoid pathway. The article can be found at; <a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3287197/pdf/gkr888.pdf"> Conrado RJ, Mori J et al (2011) </a><br />
<br />
<br><br />
<br><br />
<br />
<h1> Other collaborations </h1><br />
<br />
In addition to this we would also like to mention some fascinating requests that have been made by external sources: <br />
<ul><br />
<li> Nature reviews request to display our chromoproteins on their <a href="http://www.nature.com/nrmicro/journal/v11/n10/covers/index.html"> front page </a> for their 10 year anniversary. </li><br />
<li>National Aeronautics and Space Administrations (NASA) biotechnology department requested our chromoproteins as reporter genes for a project that is currently being worked on. </li><br />
</ul><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
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<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
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<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
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<h1 class="main-title">Collaboration</h1><br />
<br />
All of the work that was done during this project was both planned and executed by our iGEM team members. It all started of with some brainstorming sessions that began in the middle of May, although the actual lab work began in the beginning of June. <br />
<br />
<br><br />
<br><br />
<br />
During the summer we have worked very closely with the <a href="https://2013.igem.org/Team:Linkoping_Sweden"> Linköpings iGEM team 2013 </a>. Since it was their first year to participate in the competition we decided to do our best to provide them with any assistance they needed, such as providing them with parts, knowledge etc.<br />
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<br><br />
<br><br />
<br />
In addition to assisting Linköpings iGEM team we have also sent biobricks to the following iGEM teams:<br />
<br><br />
<br><br />
<br />
iGEM 2013 <a href="https://2013.igem.org/Team:UT-Tokyo"> UT-Tokyo: </a>:<br><br />
<a href="http://parts.igem.org/Part:BBa_K592010"> BBa_K592010 </a> - amilGFP, yellow chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K864401"> BBa_K864401 </a> - aeBlue, blue chromoprotein <br><br />
<br />
<br><br />
<br />
iGEM 2013 <a href="https://2013.igem.org/Team:Braunschweig"> Braunschweig: </a> <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033930">BBa_K1033930</a> - amilCP, blue/purple chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K592010"> BBa_K592010 </a> - amilGFP, yellow chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592012"> BBa_K592012 </a> - eforRed, red/pink chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592011"> BBa_K592011 </a> - cjBlue, green chromoprotein<br><br />
<a href="http://parts.igem.org/Part:BBa_K864401"> BBa_K864401 </a> - aeBlue, blue chromoprotein <br><br />
<br />
<br><br />
<br />
iGEM 2013 <a href="https://2013.igem.org/Team:Tsinghua"> Tsinghua: </a> <br><br />
<a href="http://parts.igem.org/Part:BBa_K592010"> BBa_K592010 </a> - amilGFP, yellow chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592012"> BBa_K592012 </a> - eforRed, red/pink chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592011"> BBa_K592011 </a> - cjBlue, green chromoprotein<br><br />
<a href="http://parts.igem.org/Part:BBa_K864401"> BBa_K864401 </a> - aeBlue, blue chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033930">BBa_K1033930</a> - amilCP, blue/purple chromoprotein (incl RBS) <br><br />
<br />
<br><br />
<br />
iGEM 2013 <a href="https://2013.igem.org/Team:CU-Boulder"> CU Boulder: </a> <br><br />
<a href="http://parts.igem.org/Part:BBa_K592010"> BBa_K592010 </a> - amilGFP, yellow chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592012"> BBa_K592012 </a> - eforRed, red/pink chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592011"> BBa_K592011 </a> - cjBlue, green chromoprotein<br><br />
<a href="http://parts.igem.org/Part:BBa_K864401"> BBa_K864401 </a> - aeBlue, blue chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033930">BBa_K1033930</a> - amilCP, blue/purple chromoprotein (incl RBS) <br><br />
<br />
<br><br />
<br />
iGEM 2013 <a href="https://2013.igem.org/Team:uOttawa"> uOttawa: </a> <br><br />
<a href="http://parts.igem.org/Part:BBa_K592010"> BBa_K592010 </a> - amilGFP, yellow chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592012"> BBa_K592012 </a> - eforRed, red/pink chromoprotein <br><br />
<br />
<br><br />
<br />
iGEM 2013 <a href="https://2013.igem.org/Team:HokkaidoU_Japan"> HokkaidoU: </a> <br><br />
<a href="http://parts.igem.org/Part:BBa_K592009"> BBa_K592009 </a> - amilCP, blue chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033931">BBa_K1033931</a> - amilGFP, yellow chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K592010"> BBa_K592010 </a> - amilGFP, yellow chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592012"> BBa_K592012 </a> - eforRed, red/pink chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592011"> BBa_K592011 </a> - cjBlue, green chromoprotein<br><br />
<a href="http://parts.igem.org/Part:BBa_K864401"> BBa_K864401 </a> - aeBlue, blue chromoprotein <br><br />
<br />
<br><br />
<br />
iGEM 2013 <a href="https://2013.igem.org/Team:TU_Darmstadt"> TU Darmstadt: </a> <br><br />
<a href="http://parts.igem.org/Part:BBa_K592012"> BBa_K592012 </a> - eforRed, red/pink chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K592011"> BBa_K592011 </a> - cjBlue, green chromoprotein<br><br />
<a href="http://parts.igem.org/Part:BBa_K1033929">BBa_K1033929</a> - aeBlue, blue chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033928">BBa_K1033928</a> - aeBlue, blue chromoprotein (incl RBS, J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033903"> BBa_K1033903 </a> - tsPurple, purple chromoprotein (incl RBS, J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033930">BBa_K1033930</a> - amilCP, blue/purple chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033931">BBa_K1033931</a> - amilGFP, yellow chromoprotein (incl RBS) <br><br />
<br />
<br><br />
<br />
<h1> A special thanks </h1><br />
<br />
We would also like to thank other sources that have helped us, such as the <a href="https://2010.igem.org/Team:Slovenia"> Slovenian iGEM team 2010 </a> that suppied us with the genes needed for the production of carotenoid metabolites, <a href="https://2012.igem.org/Team:TU_Munich"> TU-Munichs iGEM team 2012 </a> that provided us with the gene 4CL for the production of 4-coumaryl-CoA, last but not least we would also like to thank the School of chemical and Biomolecular Engineering, Cornell University for providing us with the genes involved in the production of the phenylpropanoid pathway. The article can be found at; <a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3287197/pdf/gkr888.pdf"> Conrado RJ, Mori J et al (2011) </a><br />
<br />
<br><br />
<br><br />
<br />
<h1> Other collaborations </h1><br />
<br />
In addition to this we would also like to mention some fascinating requests that have been made by external sources: <br />
<ul><br />
<li> Nature reviews request to display our chromoproteins on their <a href="http://www.nature.com/nrmicro/journal/v11/n10/covers/index.html"> front page </a> for their 10 year anniversary. </li><br />
<li>National Aeronautics and Space Administrations (NASA) biotechnology department requested our chromoproteins as reporter genes for a project that is currently being worked on. </li><br />
</ul><br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/notebookTeam:Uppsala/notebook2013-10-04T18:58:25Z<p>Sabrijamal: </p>
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/advisors">Advisors</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/protocols">Protocols</a></li><br />
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<h1 class="main-title">Sir Cow explains</h1><br />
<h3>This is a three step tutorial of the Uppsala Notebook</h3><br />
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<b style="color:#22409a;">1.</b> By pressing the arrows in the upper right and left corner, you can change months<br><br><br />
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<b style="color:#22409a;">2.</b> Click on the day or week number, to get the notes you desire.<br><br><br />
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<b style="color:#22409a;">3.</b> Click on the box that appear under the calendar.<br><br><br />
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<div><h3>Tips</h3> <div style="height:20px;width:45px;border: 2px solid #22409a;display:inline-table;"></div> <div style="display:inline; position:relative; top:-5px;">- Days with blue borders contains notes.</div></div><br />
Uppsala iGEM team was most active between june to september 2013<br><br><br />
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<i>Now you can Notebook-explore lika a Sir! <img src="https://static.igem.org/mediawiki/2013/b/b2/SirCow_no_talk_uppsala.png" style="width:40px;"></i><br />
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<h1 class="main-title-left">Zeaxanthin</h1><br />
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<p>Zeaxanthin is a yellow carotenoid pigment, derived from the precursor ß-carotene through hydroxylation by the enzyme ”Beta-carotene hydroxylase”. Zeaxanthin acts as an antioxidant and can be found in for example peppers, yolk and maize.<sup><a href="#l1">(1)</a></sup> According to studies zeaxanthin has positive effects on both undamaged and impaired vision and it may prevent age-related macular degeneration (AMD), an eye condition that could lead to blindness.<sup><a href="#l2"> (2)</a></sup> Furthermore studies have also indicated that zeaxanthin may have skin protective activities.<sup><a href="#l2"> (1,3)</a></sup><br />
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Except for having beneficial properties the carotenoid zeaxanthin is one of the primary precursors for the production of the <a href="https://2013.igem.org/Team:Uppsala/saffron">saffron metabolites</a> picrocrocin, crocin and safranal. The production of saffron was inspired by the work of Washington team <a href="https://2012.igem.org/Team:WashU">WashU iGEM 2012.</a> Since the gene (CrtZ) that was used for the production of zeaxanthin was a eukaryotic version from Arabidopsis Thaliana, we decided to focus on finding a suitable gene to optimize the production of the above-mentioned metabolites. When searching for an appropriate gene we came in contact with <a href="https://2010.igem.org/Team:Slovenia">Slovenia iGEM 2010</a> that had produced zeaxanthin in E.coli. The operon that was obtained contained genes that were originally from the bacteria <a href="http://parts.igem.org/Part:BBa_K323122">Pantoea Anantis.</a></p> <br />
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<a id="b1"><h1>Methods:</h1></a><br />
<p>For the characterization of zeaxanthin in E.coli DH5alpha we put together a standardized method for zeaxanthin extraction and measuring. Since zeaxanthin in its pure form is extremely expensive (~ 480 €/mg) we bought maize at our local supermarket and carried out liquid-liquid extractions using methanol and performed spectrophotometry measurements. We compared literature absorbance values of zeaxanthin to the peaks we obtained from our own measurements and continued our experiments until we believed to have an optimized extraction method.</p><br />
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<img class="zeaxanthin1" src="https://static.igem.org/mediawiki/2013/9/9d/Uppsala2013_Zeaxanthin1.png"><br />
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<i><b>Figure 1.</b> Maize bought at our local supermarket that was grinded and later used to make a standardized zeaxanthin extraction method. </i><br />
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<i><b>Figure 2.</b> Extraction experiments carried out on grinded maize using methanol.</i><br />
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<p>The operon acquired from Slovenia iGEM 2010 consists of the genes CrtE, CrtB, CrtI, CrtY and CrtZ assembled with the inducible promoter pBAD/AraC.<br />
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Since we would eventually like to produce zeaxanthin in Lactobacillus in yoghurt it would be preferable to have a constitutive promoter, such as the CP-promoters in our <a href="https://2013.igem.org/Team:Uppsala/promoters#l1">promoter library.</a> Parallel to the characterization of zeaxanthin in E.coli we therefore attempted to remove the pBAD/AraC promoter from the zeaxanthin operon.</p><br />
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<img class="methods-plasmid" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Zeaxanthin_Plasmid_and_molecules.png"><br />
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<h1>Results:</h1><br />
<h3>Spectrophotometry</h3><br />
<p>Production of zeaxanthin in E.coli DH5alpha was detected trough spectrophotometry measurements performed after liquid-liquid extractions using methanol. The resulting absorbance curve was compared to the standardized curve. In both graphs you can find the two peaks corresponding to zeaxanthin.</p><br />
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<img class="zeaxanthin3" src="https://static.igem.org/mediawiki/2013/3/30/Uppsala2013_zeaxanthin2.png"><br />
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<i><b>Figure 3.</b> E.coli culture believed to have zeaxanthin production.</i><br />
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<i><b>Figure 4.</b> A liquid-liquid extraction was carried out on the E.coli culture containing the plasmid above. Methanol was used as an organic solvent.</i><br />
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<img class="zeaxanthin5" src="https://static.igem.org/mediawiki/2013/0/04/Uppsala2013_Zeaxanthin5.jpg"><br />
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<i><b>Figure 5.</b> Spectrophotometry of zeaxanthin extraction from maize resulted in the graph above. The two peaks are characteristic for zeaxanthin.</i><br />
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<i><b>Figure 6.</b> Spectrophotometry of an E.coli culture containing the plasmid with the zeaxanthin operon. The characteristic peaks of zeaxanthin are present, indicating that zeaxanthin was present in the sample.</i><br />
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<p>Spectrophotometry measurements were also done on unmodified E.coli, as a negative control. These measurements did not result in the peaks that are characteristic for zeaxanthin. </p><br />
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<img class="zeaxanthin7" src="https://static.igem.org/mediawiki/2013/d/d9/Uppsala2013_Zeaxanthin7.jpg"><br />
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<i class="fig-text7"><b>Figure 7.</b> Spectrophotometry of unmodified E.coli did not result<br> in the peaks characteristic for zeaxanthin. </i><br />
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<h1> References </h1><br />
[1]. Baumann, Leslie S. “Zeaxanthin.(Cosmeceutical critique)” Skin & Allergy News. ISSN: 0037-6337, Volume 40, Issue 5. 2009<br />
<br><br />
[2]. Loskutova E et al, “Macular Pigment and its Contribution to Vision” Nutrients ISSN 2072-6643, 2013<br />
<br><br />
[3]. ”Protection against sunburn and skin problems with orally-ingested high-dosage zeaxanthin”<br />
<a href=”http://apps.webofknowledge.com.ezproxy.its.uu.se/full_record.do?product=UA&search_mode=GeneralSearch&qid=1&SID=S2MoOQiIQ1bvPdZp3S6&page=1&doc=10”> http://apps.webofknowledge.com.ezproxy.its.uu.se/full_record.doproduct=UA&search_mode=GeneralSearch&qid=1&SID=S2MoOQiIQ1bvPdZp3S6&page=1&<br />
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<h1 class="main-title-left">Zeaxanthin</h1><br />
<img class="metabolic-pic" src="https://static.igem.org/mediawiki/2013/e/e5/Uppsala2013_Zeaxantin2symbol.png"><br />
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<p>Zeaxanthin is a yellow carotenoid pigment, derived from the precursor ß-carotene through hydroxylation by the enzyme ”Beta-carotene hydroxylase”. Zeaxanthin acts as an antioxidant and can be found in for example peppers, yolk and maize.<sup><a href="#l1">(1)</a></sup> According to studies zeaxanthin has positive effects on both undamaged and impaired vision and it may prevent age-related macular degeneration (AMD), an eye condition that could lead to blindness.<sup><a href="#l2"> (2)</a></sup> Furthermore studies have also indicated that zeaxanthin may have skin protective activities.<sup><a href="#l2"> (1,3)</a></sup><br />
<br><br><br />
<br />
Except for having beneficial properties the carotenoid zeaxanthin is one of the primary precursors for the production of the <a href="https://2013.igem.org/Team:Uppsala/saffron">saffron metabolites</a> picrocrocin, crocin and safranal. The production of saffron was inspired by the work of Washington team <a href="https://2012.igem.org/Team:WashU">WashU iGEM 2012.</a> Since the gene (CrtZ) that was used for the production of zeaxanthin was a eukaryotic version from Arabidopsis Thaliana, we decided to focus on finding a suitable gene to optimize the production of the above-mentioned metabolites. When searching for an appropriate gene we came in contact with <a href="https://2010.igem.org/Team:Slovenia">Slovenia iGEM 2010</a> that had produced zeaxanthin in E.coli. The operon that was obtained contained genes that were originally from the bacteria <a href="http://parts.igem.org/Part:BBa_K323122">Pantoea Anantis.</a></p> <br />
<br />
<a id="b1"><h1>Methods:</h1></a><br />
<p>For the characterization of zeaxanthin in E.coli DH5alpha we put together a standardized method for zeaxanthin extraction and measuring. Since zeaxanthin in its pure form is extremely expensive (~ 480 €/mg) we bought maize at our local supermarket and carried out liquid-liquid extractions using methanol and performed spectrophotometry measurements. We compared literature absorbance values of zeaxanthin to the peaks we obtained from our own measurements and continued our experiments until we believed to have an optimized extraction method.</p><br />
<br />
<img class="zeaxanthin1" src="https://static.igem.org/mediawiki/2013/9/9d/Uppsala2013_Zeaxanthin1.png"><br />
<br />
<i><b>Figure 1.</b> Maize bought at our local supermarket that was grinded and later used to make a standardized zeaxanthin extraction method. </i><br />
<br><br />
<i><b>Figure 2.</b> Extraction experiments carried out on grinded maize using methanol.</i><br />
<br><br> <br />
<br />
<p>The operon acquired from Slovenia iGEM 2010 consists of the genes CrtE, CrtB, CrtI, CrtY and CrtZ assembled with the inducible promoter pBAD/AraC.<br />
<br><br><br />
<br />
Since we would eventually like to produce zeaxanthin in Lactobacillus in yoghurt it would be preferable to have a constitutive promoter, such as the CP-promoters in our <a href="https://2013.igem.org/Team:Uppsala/promoters#l1">promoter library.</a> Parallel to the characterization of zeaxanthin in E.coli we therefore attempted to remove the pBAD/AraC promoter from the zeaxanthin operon.</p><br />
<br />
<img class="methods-plasmid" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Zeaxanthin_Plasmid_and_molecules.png"><br />
<br />
<h1>Results:</h1><br />
<h3>Spectrophotometry</h3><br />
<p>Production of zeaxanthin in E.coli DH5alpha was detected trough spectrophotometry measurements performed after liquid-liquid extractions using methanol. The resulting absorbance curve was compared to the standardized curve. In both graphs you can find the two peaks corresponding to zeaxanthin.</p><br />
<br />
<img class="zeaxanthin3" src="https://static.igem.org/mediawiki/2013/3/30/Uppsala2013_zeaxanthin2.png"><br />
<br />
<br />
<i><b>Figure 3.</b> E.coli culture believed to have zeaxanthin production.</i><br />
<br><br />
<i><b>Figure 4.</b> A liquid-liquid extraction was carried out on the E.coli culture containing the plasmid above. Methanol was used as an organic solvent.</i><br />
<br />
<div id="pic-box3"><br />
<img class="zeaxanthin5" src="https://static.igem.org/mediawiki/2013/0/04/Uppsala2013_Zeaxanthin5.jpg"><br />
<img class="zeaxanthin6" src="https://static.igem.org/mediawiki/2013/0/0e/Uppsala2013_Zeaxanthin6.jpg"><br />
</div> <br />
<br />
<div id="clear"></div><br />
<br />
<i><b>Figure 5.</b> Spectrophotometry of zeaxanthin extraction from maize resulted in the graph above. The two peaks are characteristic for zeaxanthin.</i><br />
<br><br />
<i><b>Figure 6.</b> Spectrophotometry of an E.coli culture containing the plasmid with the zeaxanthin operon. The characteristic peaks of zeaxanthin are present, indicating that zeaxanthin was present in the sample.</i><br />
<br />
<br />
<br><br><br />
<p>Spectrophotometry measurements were also done on unmodified E.coli, as a negative control. These measurements did not result in the peaks that are characteristic for zeaxanthin. </p><br />
<br />
<img class="zeaxanthin7" src="https://static.igem.org/mediawiki/2013/d/d9/Uppsala2013_Zeaxanthin7.jpg"><br />
<br />
<i class="fig-text7"><b>Figure 7.</b> Spectrophotometry of unmodified E.coli did not result<br> in the peaks characteristic for zeaxanthin. </i><br />
<br />
<div id="clear"></div><br />
<br />
<h1> References </h1><br />
[1]. Baumann, Leslie S. “Zeaxanthin.(Cosmeceutical critique)” Skin & Allergy News. ISSN: 0037-6337, Volume 40, Issue 5. 2009<br />
<br><br />
[2]. Loskutova E et al, “Macular Pigment and its Contribution to Vision” Nutrients ISSN 2072-6643, 2013<br />
<br><br />
[3]. ”Protection against sunburn and skin problems with orally-ingested high-dosage zeaxanthin”<br />
<a href=”http://apps.webofknowledge.com.ezproxy.its.uu.se/full_record.do?product=UA&search_mode=GeneralSearch&qid=1&SID=S2MoOQiIQ1bvPdZp3S6&page=1&doc=10”> http://apps.webofknowledge.com.ezproxy.its.uu.se/full_record.doproduct=UA&search_mode=GeneralSearch&qid=1&SID=S2MoOQiIQ1bvPdZp3S6&page=1&doc=10 </a> <br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/zeaxanthinTeam:Uppsala/zeaxanthin2013-10-04T17:55:17Z<p>Sabrijamal: </p>
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<h1 class="main-title-left">Zeaxanthin</h1><br />
<img class="metabolic-pic" src="https://static.igem.org/mediawiki/2013/e/e5/Uppsala2013_Zeaxantin2symbol.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>Zeaxanthin is a yellow carotenoid pigment, derived from the precursor ß-carotene through hydroxylation by the enzyme ”Beta-carotene hydroxylase”. Zeaxanthin acts as an antioxidant and can be found in for example peppers, yolk and maize.<sup><a href="#l1">(1)</a></sup> According to studies zeaxanthin has positive effects on both undamaged and impaired vision and it may prevent age-related macular degeneration (AMD), an eye condition that could lead to blindness.<sup><a href="#l2"> (2)</a></sup> Furthermore studies have also indicated that zeaxanthin may have skin protective activities.<sup><a href="#l2"> (1,3)</a></sup><br />
<br><br><br />
<br />
Except for having beneficial properties the carotenoid zeaxanthin is one of the primary precursors for the production of the <a href="https://2013.igem.org/Team:Uppsala/saffron">saffron metabolites</a> picrocrocin, crocin and safranal. The production of saffron was inspired by the work of Washington team <a href="https://2012.igem.org/Team:WashU">WashU iGEM 2012.</a> Since the gene (CrtZ) that was used for the production of zeaxanthin was a eukaryotic version from Arabidopsis Thaliana, we decided to focus on finding a suitable gene to optimize the production of the above-mentioned metabolites. When searching for an appropriate gene we came in contact with <a href="https://2010.igem.org/Team:Slovenia">Slovenia iGEM 2010</a> that had produced zeaxanthin in E.coli. The operon that was obtained contained genes that were originally from the bacteria <a href="http://parts.igem.org/Part:BBa_K323122">Pantoea Anantis.</a></p> <br />
<br />
<a id="b1"><h1>Methods:</h1></a><br />
<p>For the characterization of zeaxanthin in E.coli DH5alpha we put together a standardized method for zeaxanthin extraction and measuring. Since zeaxanthin in its pure form is extremely expensive (~ 480 €/mg) we bought maize at our local supermarket and carried out liquid-liquid extractions using methanol and performed spectrophotometry measurements. We compared literature absorbance values of zeaxanthin to the peaks we obtained from our own measurements and continued our experiments until we believed to have an optimized extraction method.</p><br />
<br />
<img class="zeaxanthin1" src="https://static.igem.org/mediawiki/2013/9/9d/Uppsala2013_Zeaxanthin1.png"><br />
<br />
<i><b>Figure 1.</b> Maize bought at our local supermarket that was grinded and later used to make a standardized zeaxanthin extraction method. </i><br />
<br><br />
<i><b>Figure 2.</b> Extraction experiments carried out on grinded maize using methanol.</i><br />
<br><br> <br />
<br />
<p>The operon acquired from Slovenia iGEM 2010 consists of the genes CrtE, CrtB, CrtI, CrtY and CrtZ assembled with the inducible promoter pBAD/AraC.<br />
<br><br><br />
<br />
Since we would eventually like to produce zeaxanthin in Lactobacillus in yoghurt it would be preferable to have a constitutive promoter, such as the CP-promoters in our <a href="https://2013.igem.org/Team:Uppsala/promoters#l1">promoter library.</a> Parallel to the characterization of zeaxanthin in E.coli we therefore attempted to remove the pBAD/AraC promoter from the zeaxanthin operon.</p><br />
<br />
<img class="methods-plasmid" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Zeaxanthin_Plasmid_and_molecules.png"><br />
<br />
<h1>Results:</h1><br />
<h3>Spectrophotometry</h3><br />
<p>Production of zeaxanthin in E.coli DH5alpha was detected trough spectrophotometry measurements performed after liquid-liquid extractions using methanol. The resulting absorbance curve was compared to the standardized curve. In both graphs you can find the two peaks corresponding to zeaxanthin.</p><br />
<br />
<img class="zeaxanthin3" src="https://static.igem.org/mediawiki/2013/3/30/Uppsala2013_zeaxanthin2.png"><br />
<br />
<br />
<i><b>Figure 3.</b> E.coli culture believed to have zeaxanthin production.</i><br />
<br><br />
<i><b>Figure 4.</b> A liquid-liquid extraction was carried out on the E.coli culture containing the plasmid above. Methanol was used as an organic solvent.</i><br />
<br />
<div id="pic-box3"><br />
<img class="zeaxanthin5" src="https://static.igem.org/mediawiki/2013/0/04/Uppsala2013_Zeaxanthin5.jpg"><br />
<img class="zeaxanthin6" src="https://static.igem.org/mediawiki/2013/0/0e/Uppsala2013_Zeaxanthin6.jpg"><br />
</div> <br />
<br />
<div id="clear"></div><br />
<br />
<i><b>Figure 5.</b> Spectrophotometry of zeaxanthin extraction from maize resulted in the graph above. The two peaks are characteristic for zeaxanthin.</i><br />
<br><br />
<i><b>Figure 6.</b> Spectrophotometry of an E.coli culture containing the plasmid with the zeaxanthin operon. The characteristic peaks of zeaxanthin are present, indicating that zeaxanthin was present in the sample.</i><br />
<br />
<br />
<br><br><br />
<p>Spectrophotometry measurements were also done on unmodified E.coli, as a negative control. These measurements did not result in the peaks that are characteristic for zeaxanthin. </p><br />
<br />
<img class="zeaxanthin7" src="https://static.igem.org/mediawiki/2013/d/d9/Uppsala2013_Zeaxanthin7.jpg"><br />
<br />
<i class="fig-text7"><b>Figure 7.</b> Spectrophotometry of unmodified E.coli did not result<br> in the peaks characteristic for zeaxanthin. </i><br />
<br />
<div id="clear"></div><br />
<br />
<h1> References </h1><br />
[1]. Baumann, Leslie S. “Zeaxanthin.(Cosmeceutical critique)” Skin & Allergy News. ISSN: 0037-6337, Volume 40, Issue 5. 2009<br />
<br><br />
[2]. Loskutova E et al, “Macular Pigment and its Contribution to Vision” Nutrients ISSN 2072-6643, 2013<br />
<br><br />
[3]. ”Protection against sunburn and skin problems with orally-ingested high-dosage zeaxanthin”<br />
<a href=”http://apps.webofknowledge.com.ezproxy.its.uu.se/full_record.do?product=UA&search_mode=GeneralSearch&qid=1&SID=S2MoOQiIQ1bvPdZp3S6&page=1&doc=10”> http://apps.webofknowledge.com.ezproxy.its.uu.se/full_record.do?product=UA&search_mode=GeneralSearch&qid=1&SID=S2MoOQiIQ1bvPdZp3S6&page=1&doc=10 </a> <br />
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<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
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<h1 class="main-title-left">Zeaxanthin</h1><br />
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<p>Zeaxanthin is a yellow carotenoid pigment, derived from the precursor ß-carotene through hydroxylation by the enzyme ”Beta-carotene hydroxylase”. Zeaxanthin acts as an antioxidant and can be found in for example peppers, yolk and maize.<sup><a href="#l1">(1)</a></sup> According to studies zeaxanthin has positive effects on both undamaged and impaired vision and it may prevent age-related macular degeneration (AMD), an eye condition that could lead to blindness.<sup><a href="#l2"> (2)</a></sup> Furthermore studies have also indicated that zeaxanthin may have skin protective activities.<sup><a href="#l2"> (1,3)</a></sup><br />
<br><br><br />
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Except for having beneficial properties the carotenoid zeaxanthin is one of the primary precursors for the production of the <a href="https://2013.igem.org/Team:Uppsala/saffron">saffron metabolites</a> picrocrocin, crocin and safranal. The production of saffron was inspired by the work of Washington team <a href="https://2012.igem.org/Team:WashU">WashU iGEM 2012.</a> Since the gene (CrtZ) that was used for the production of zeaxanthin was a eukaryotic version from Arabidopsis Thaliana, we decided to focus on finding a suitable gene to optimize the production of the above-mentioned metabolites. When searching for an appropriate gene we came in contact with <a href="https://2010.igem.org/Team:Slovenia">Slovenia iGEM 2010</a> that had produced zeaxanthin in E.coli. The operon that was obtained contained genes that were originally from the bacteria <a href="http://parts.igem.org/Part:BBa_K323122">Pantoea Anantis.</a></p> <br />
<br />
<a id="b1"><h1>Methods:</h1></a><br />
<p>For the characterization of zeaxanthin in E.coli DH5alpha we put together a standardized method for zeaxanthin extraction and measuring. Since zeaxanthin in its pure form is extremely expensive (~ 480 €/mg) we bought maize at our local supermarket and carried out liquid-liquid extractions using methanol and performed spectrophotometry measurements. We compared literature absorbance values of zeaxanthin to the peaks we obtained from our own measurements and continued our experiments until we believed to have an optimized extraction method.</p><br />
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<img class="zeaxanthin1" src="https://static.igem.org/mediawiki/2013/9/9d/Uppsala2013_Zeaxanthin1.png"><br />
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<i><b>Figure 1.</b> Maize bought at our local supermarket that was grinded and later used to make a standardized zeaxanthin extraction method. </i><br />
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<i><b>Figure 2.</b> Extraction experiments carried out on grinded maize using methanol.</i><br />
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<p>The operon acquired from Slovenia iGEM 2010 consists of the genes CrtE, CrtB, CrtI, CrtY and CrtZ assembled with the inducible promoter pBAD/AraC.<br />
<br><br><br />
<br />
Since we would eventually like to produce zeaxanthin in Lactobacillus in yoghurt it would be preferable to have a constitutive promoter, such as the CP-promoters in our <a href="https://2013.igem.org/Team:Uppsala/promoters#l1">promoter library.</a> Parallel to the characterization of zeaxanthin in E.coli we therefore attempted to remove the pBAD/AraC promoter from the zeaxanthin operon.</p><br />
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<img class="methods-plasmid" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Zeaxanthin_Plasmid_and_molecules.png"><br />
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<h1>Results:</h1><br />
<h3>Spectrophotometry</h3><br />
<p>Production of zeaxanthin in E.coli DH5alpha was detected trough spectrophotometry measurements performed after liquid-liquid extractions using methanol. The resulting absorbance curve was compared to the standardized curve. In both graphs you can find the two peaks corresponding to zeaxanthin.</p><br />
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<img class="zeaxanthin3" src="https://static.igem.org/mediawiki/2013/3/30/Uppsala2013_zeaxanthin2.png"><br />
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<i><b>Figure 3.</b> E.coli culture believed to have zeaxanthin production.</i><br />
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<i><b>Figure 4.</b> A liquid-liquid extraction was carried out on the E.coli culture containing the plasmid above. Methanol was used as an organic solvent.</i><br />
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<img class="zeaxanthin5" src="https://static.igem.org/mediawiki/2013/0/04/Uppsala2013_Zeaxanthin5.jpg"><br />
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<i><b>Figure 5.</b> Spectrophotometry of zeaxanthin extraction from maize resulted in the graph above. The two peaks are characteristic for zeaxanthin.</i><br />
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<i><b>Figure 6.</b> Spectrophotometry of an E.coli culture containing the plasmid with the zeaxanthin operon. The characteristic peaks of zeaxanthin are present, indicating that zeaxanthin was present in the sample.</i><br />
<br />
<br />
<br><br><br />
<p>Spectrophotometry measurements were also done on unmodified E.coli, as a negative control. These measurements did not result in the peaks that are characteristic for zeaxanthin. </p><br />
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<img class="zeaxanthin7" src="https://static.igem.org/mediawiki/2013/d/d9/Uppsala2013_Zeaxanthin7.jpg"><br />
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<i class="fig-text7"><b>Figure 7.</b> Spectrophotometry of unmodified E.coli did not result<br> in the peaks characteristic for zeaxanthin. </i><br />
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<h1> References </h1><br />
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<h1 class="main-title-left">Lycopene</h1><br />
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<p class="text-left">Lycopene is a carotenoid that primarily derives from the xanthophyll group. Lycopene is an antioxidant that can be found in both vegetables and fruits although it is mostly known for being responsible for the characteristic red color in tomatoes. Several studies indicate that lycopene can decrease the risk of cancer and heart diseases. <sup> <a href="#refpoint"> [1] </a> </sup>Because of the antioxidant capacities as well as being an important part of the carotenoid pathway makes it an interesting part to study further.</p><br />
<br />
<img class="molecule-pic" src="https://static.igem.org/mediawiki/2013/8/89/Uppsala2013_lycopene-molecule.png"><br />
<br />
<a id="b1"></a><h1>Methods</h1><br />
<p>From Slovenia iGEM team 2010 we got an operon containing CrtE, CrtB, CrtI, zinc fingers and linkers were assembled with the inducible promoter pBAD/AraC. In our project the zinc fingers were not put into a lot of consideration. The precursor responsible for lycopene production is phytoene. When phytoene is present together with the enzyme phytoene synthase which is translated by CrtI which was isolated from the operon a catalytic reaction occurs.<br />
A standardize test for lycopene was put together by extraction of tomatoes. The tomatoes were boiled in ethyl-acetate to separate. DH5-alpha bacteria which should produce lycopene with the help of our plasmid were separated in a liquid-liquid separation with ethyl-acetate.</p><br />
<br />
<img class="methods-pic" src="https://static.igem.org/mediawiki/2013/9/9d/Uppsala2013_methods_Lycopene.png"><br />
<br />
<h1>Results</h1><br />
<br />
A test for lycopene was put together by extraction of lycopene from tomatoes. The tomatoes were boiled in ethyl-acetate and centrifuged to separate the different compounds and in the case with the DH5-alpha bacteria cultures we also used liquid-liquid extractions. <br />
<br />
<br><br />
<br><br />
<br />
<div id="lycopene_extraction"> <img class="lycopene_extraction" src="https://static.igem.org/mediawiki/2013/2/29/Uppsala2013_lycopene_crush.png"> <br />
<i> Figure 1, Freshly squeezed tomatoes purchased at our local store <br><br />
Figure 2, Squeezed tomatoes diluted in ethyl acetetate used for further <br><br />
extraction purposes </i><br />
<br><br />
<br><br />
<br />
</div> <br />
<br />
<p>The pellet of the bacteria test showed a red pellet which would indicate the existence of lycopene. The tomato extraction test showed a red liquid which is a strong indication regarding the existence of lycopene. The tomato extraction was set as the standard and tries measuring the concentration in bacteria's producing lycopene was used. Unfortunately the concentration was too small to be detected clearly as seen below in the graphs.</p> <br />
<br />
<br><br />
<h2>Spectrophotometry</h2><br />
<table><br />
<tr><td class="pic_col_results_pc"><img class="results_pic_pc" src="https://static.igem.org/mediawiki/2013/a/af/Lycopene2.png"></td><td class="fig-text"><i><b>Figure 3</b> Lycopene test from tomato, all 3 characteristic peaks are at correct wavelength and strength, we will use this as out standard for lycopene from bacteria cultures.</i> </td></tr><br />
<tr><td class="pic_col_results_pc"><img class="results_pic_pc" src="https://static.igem.org/mediawiki/2013/6/60/LycopeneNC.png"></td><td class="fig-text"><i><b>Figure 4</b> Negative control in ethyl acetate.</i></td></tr><br />
<tr><td class="pic_col_results_pc"><img class="results_pic_pc" src="https://static.igem.org/mediawiki/2013/c/cc/Lycopene3.png"></td><td class="fig-text"><i><b>Figure 5</b> Extraction try for lycopene from bacteria culture, due to lack of time we didn´t do more than 1 try to extract it which sadly didn´t give any good result.</i> </td></tr><br />
</table><br />
<br />
<h1> References </h1><br />
1. Rao and Agarwal S - Role of Antioxidant Lycopene in Cancer and Heart Disease, J Am Coll Nutr. 2000 Oct;19(5):563-9.<br />
<br />
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<h1 class="main-title-left">Lycopene</h1><br />
<br />
<img class="metabolic-pic" src="https://static.igem.org/mediawiki/2013/8/8f/Uppsala2013_LycopeneSymbol.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p class="text-left">Lycopene is a carotenoid that primarily derives from the xanthophyll group. Lycopene is an antioxidant that can be found in both vegetables and fruits although it is mostly known for being responsible for the characteristic red color in tomatoes. Several studies indicate that lycopene can decrease the risk of cancer and heart diseases. <sup> [1] </sup>Because of the antioxidant capacities as well as being an important part of the carotenoid pathway makes it an interesting part to study further.</p><br />
<br />
<img class="molecule-pic" src="https://static.igem.org/mediawiki/2013/8/89/Uppsala2013_lycopene-molecule.png"><br />
<br />
<a id="b1"></a><h1>Methods</h1><br />
<p>From Slovenia iGEM team 2010 we got an operon containing CrtE, CrtB, CrtI, zinc fingers and linkers were assembled with the inducible promoter pBAD/AraC. In our project the zinc fingers were not put into a lot of consideration. The precursor responsible for lycopene production is phytoene. When phytoene is present together with the enzyme phytoene synthase which is translated by CrtI which was isolated from the operon a catalytic reaction occurs.<br />
A standardize test for lycopene was put together by extraction of tomatoes. The tomatoes were boiled in ethyl-acetate to separate. DH5-alpha bacteria which should produce lycopene with the help of our plasmid were separated in a liquid-liquid separation with ethyl-acetate.</p><br />
<br />
<img class="methods-pic" src="https://static.igem.org/mediawiki/2013/9/9d/Uppsala2013_methods_Lycopene.png"><br />
<br />
<h1>Results</h1><br />
<br />
A test for lycopene was put together by extraction of lycopene from tomatoes. The tomatoes were boiled in ethyl-acetate and centrifuged to separate the different compounds and in the case with the DH5-alpha bacteria cultures we also used liquid-liquid extractions. <br />
<br />
<br><br />
<br><br />
<br />
<div id="lycopene_extraction"> <img class="lycopene_extraction" src="https://static.igem.org/mediawiki/2013/2/29/Uppsala2013_lycopene_crush.png"> <br />
<i> Figure 1, Freshly squeezed tomatoes purchased at our local store <br><br />
Figure 2, Squeezed tomatoes diluted in ethyl acetetate used for further <br><br />
extraction purposes </i><br />
<br><br />
<br><br />
<br />
</div> <br />
<br />
<p>The pellet of the bacteria test showed a red pellet which would indicate the existence of lycopene. The tomato extraction test showed a red liquid which is a strong indication regarding the existence of lycopene. The tomato extraction was set as the standard and tries measuring the concentration in bacteria's producing lycopene was used. Unfortunately the concentration was too small to be detected clearly as seen below in the graphs.</p> <br />
<br />
<br><br />
<h2>Spectrophotometry</h2><br />
<table><br />
<tr><td class="pic_col_results_pc"><img class="results_pic_pc" src="https://static.igem.org/mediawiki/2013/a/af/Lycopene2.png"></td><td class="fig-text"><i><b>Figure 3</b> Lycopene test from tomato, all 3 characteristic peaks are at correct wavelength and strength, we will use this as out standard for lycopene from bacteria cultures.</i> </td></tr><br />
<tr><td class="pic_col_results_pc"><img class="results_pic_pc" src="https://static.igem.org/mediawiki/2013/6/60/LycopeneNC.png"></td><td class="fig-text"><i><b>Figure 4</b> Negative control in ethyl acetate.</i></td></tr><br />
<tr><td class="pic_col_results_pc"><img class="results_pic_pc" src="https://static.igem.org/mediawiki/2013/c/cc/Lycopene3.png"></td><td class="fig-text"><i><b>Figure 5</b> Extraction try for lycopene from bacteria culture, due to lack of time we didn´t do more than 1 try to extract it which sadly didn´t give any good result.</i> </td></tr><br />
</table><br />
<br />
<h1> References </h1> <br />
1. Rao and Agarwal S - Role of Antioxidant Lycopene in Cancer and Heart Disease, J Am Coll Nutr. 2000 Oct;19(5):563-9.<br />
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<h1 class="main-title-left">Lycopene</h1><br />
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<p class="text-left">Lycopene is a carotenoid that primarily derives from the xanthophyll group. Lycopene is an antioxidant that can be found in both vegetables and fruits although it is mostly known for being responsible for the characteristic red color in tomatoes. Several studies indicate that lycopene can decrease the risk of cancer and heart diseases (1). Because of the antioxidant capacities as well as being an important part of the carotenoid pathway makes it an interesting part to study further.</p><br />
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<img class="molecule-pic" src="https://static.igem.org/mediawiki/2013/8/89/Uppsala2013_lycopene-molecule.png"><br />
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<a id="b1"></a><h1>Methods</h1><br />
<p>From Slovenia iGEM team 2010 we got an operon containing CrtE, CrtB, CrtI, zinc fingers and linkers were assembled with the inducible promoter pBAD/AraC. In our project the zinc fingers were not put into a lot of consideration. The precursor responsible for lycopene production is phytoene. When phytoene is present together with the enzyme phytoene synthase which is translated by CrtI which was isolated from the operon a catalytic reaction occurs.<br />
A standardize test for lycopene was put together by extraction of tomatoes. The tomatoes were boiled in ethyl-acetate to separate. DH5-alpha bacteria which should produce lycopene with the help of our plasmid were separated in a liquid-liquid separation with ethyl-acetate.</p><br />
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<img class="methods-pic" src="https://static.igem.org/mediawiki/2013/9/9d/Uppsala2013_methods_Lycopene.png"><br />
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<h1>Results</h1><br />
<br />
A test for lycopene was put together by extraction of lycopene from tomatoes. The tomatoes were boiled in ethyl-acetate and centrifuged to separate the different compounds and in the case with the DH5-alpha bacteria cultures we also used liquid-liquid extractions. <br />
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<div id="lycopene_extraction"> <img class="lycopene_extraction" src="https://static.igem.org/mediawiki/2013/2/29/Uppsala2013_lycopene_crush.png"> <br />
<i> Figure 1, Freshly squeezed tomatoes purchased at our local store <br><br />
Figure 2, Squeezed tomatoes diluted in ethyl acetetate used for further <br><br />
extraction purposes </i><br />
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<p>The pellet of the bacteria test showed a red pellet which would indicate the existence of lycopene. The tomato extraction test showed a red liquid which is a strong indication regarding the existence of lycopene. The tomato extraction was set as the standard and tries measuring the concentration in bacteria's producing lycopene was used. Unfortunately the concentration was too small to be detected clearly as seen below in the graphs.</p> <br />
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<br><br />
<h2>Spectrophotometry</h2><br />
<table><br />
<tr><td class="pic_col_results_pc"><img class="results_pic_pc" src="https://static.igem.org/mediawiki/2013/a/af/Lycopene2.png"></td><td class="fig-text"><i><b>Figure 3</b> Lycopene test from tomato, all 3 characteristic peaks are at correct wavelength and strength, we will use this as out standard for lycopene from bacteria cultures.</i> </td></tr><br />
<tr><td class="pic_col_results_pc"><img class="results_pic_pc" src="https://static.igem.org/mediawiki/2013/6/60/LycopeneNC.png"></td><td class="fig-text"><i><b>Figure 4</b> Negative control in ethyl acetate.</i></td></tr><br />
<tr><td class="pic_col_results_pc"><img class="results_pic_pc" src="https://static.igem.org/mediawiki/2013/c/cc/Lycopene3.png"></td><td class="fig-text"><i><b>Figure 5</b> Extraction try for lycopene from bacteria culture, due to lack of time we didn´t do more than 1 try to extract it which sadly didn´t give any good result.</i> </td></tr><br />
</table><br />
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<h1> References </h1> <br />
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<br><br />
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1. Rao and Agarwal S - Role of Antioxidant Lycopene in Cancer and Heart Disease, J Am Coll Nutr. 2000 Oct;19(5):563-9.<br />
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<h1 class="main-title-left">β-carotene</h1><br />
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<img class="metabolic-pic" src="https://static.igem.org/mediawiki/2013/0/00/Uppsala2013_beta-caroteneSymbol.png"><br />
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<p>β-carotene is a common carotenoid that derives from the xanthophyll group known for its characteristic orange color. β-carotene can be found in fruits and vegetables, for example carrots. β-carotene is a provitamin to vitamin A, which means that during the right circumstances β-carotene can be catalyzed further to produce vitamin A. Vitamin A has several important health aspects which includes skin, mucus membrane and most common to people a good effect on eyes and vision. Studies have shown that the carotenoid can prevent aging of the brain and stimulate it to keep its capability of memory when a high ascorbic acid concentration also is present <sup><a href="#l1">(1)</a></sup>. The health aspects were an important part of our choice to produce β-carotene.</p><br />
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<img class="molecule" src="https://static.igem.org/mediawiki/2013/f/f4/Uppsala2013_Beta-carotene-molecule.png"><br />
<br />
<a id="b1"></a><h1>Methods</h1><br />
<p>The gene CrtY that is responsible for translating the protein lycopene cyclase was obtained from the zeaxanthin operon provided by Slovenia iGEM team 2010. Production of β-carotene is initiated through the catalytic reaction done by the enzyme lycopene cyclase which uses the precursor <a href="https://2013.igem.org/Team:Uppsala/lycopene">lycopene.</a></p><br />
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<img class="methods-pic" src="https://static.igem.org/mediawiki/2013/d/d2/Uppsala2013_Beta-caroten-plasmid.png"><br />
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<h1>Results </h1><br />
<p>Beta carotene production was proven through the production of our operon containing <a href="https://2013.igem.org/Team:Uppsala/zeaxantin">zeaxanthin.</a> Since production of zeaxanthin relies on beta carotene as a precursor we could simultaneously prove that we successfully had produced both zeaxanthin and beta carotene through liquid-liquid separation and spectrophotometry <a href="https://2013.igem.org/Team:Uppsala/zeaxantin">(read spectrophotometry of zeaxanthin for more results)</a></p><br />
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<h1>Results (Bio-bricks)</h1><br />
<p>We realized that there were a lot of components in the carotenoid pathway that we were in need of in the beginning of the project that were missing in the registry. Therefore after obtaining the zeaxanthin operon from Slovenias iGEM team 2010 we directly proceeded with isolating of the genes as basic components in order to send them to the registry. </p> <br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
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</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
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<h1 class="main-title"> iGEM Uppsala chromoprotein collection </h1><br />
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<h1> Bringing color to the world of synthetic biology since 2011 </h1><br />
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<div id="chromoprotein_world"> <img class="chromoprotein_world" src="https://static.igem.org/mediawiki/2013/8/8d/Uppsala2013_Kromokarta.png"> </div> <br />
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Something we have come to realise is that sometimes the most useful parts are also the simplest. IGEM Uppsala first began synthesising several chromoproteins from sea anemones and corals back in 2011 and transforming them into E. coli. The result was the growth of strikingly colourful bacterial colonies. Work on the chromoproteins continued during the 2012 project, however because of a mix up nothing was sent to the registry.<br />
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Since then we have expanded the collection further and the interest in them has exploded. In this year alone around 10 teams have contacted us requesting our chromoproteins. They have been featured on the parts registry front page, published on the front cover of Nature reviews microbiology 10 year anniversary issue and been requested by NASA for possible future space applications. <br />
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<div id="chromoprotein_n_reviews"> <img class="chromoprotein_n_reviews" src="https://static.igem.org/mediawiki/2013/4/4b/Uppsala2013_Naturereviews.png"> </div><br />
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This is why iGEM Uppsala is proud to present our complete chromoprotein collection. We have sent the chromoproteins to the registry, assembled both with and without RBS and together with a range of popular promoters to make them as available and easy to use as possible. Our hope is that by providing a comprehensive list of different chromoproteins to the registry that others will be able to use them and continue to find new purposes for them. Below you can see some of our chromoproteins previewed.<br />
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<div id="chromoprotein_all"> <img class="chromoprotein_all" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_chromoproteins.JPG"></div><br />
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<h1> Complete chromoprotein collection </h1><br />
<a href="http://parts.igem.org/Part:BBa_K1033900">BBa_K1033900</a> - meffBlue, blue chromoprotein (incl RBS, J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033901">BBa_K1033901</a> - meffBlue, blue chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033902">BBa_K1033902</a> - meffBlue, blue chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033903">BBa_K1033903</a> - tsPurple, purple chromoprotein (incl RBS, J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033904">BBa_K1033904</a> - tsPurple, purple chromoprotein (incl RBS, J23106) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033905">BBa_K1033905</a> - tsPurple, purple chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033906">BBa_K1033906</a> - tsPurple, purple chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033907">BBa_K1033907</a> - fwYellow, yellow chromoprotein (incl RBS, J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033908">BBa_K1033908</a> - fwYellow, yellow chromoprotein (incl RBS, J23106) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033909">BBa_K1033909</a> - fwYellow, yellow chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033910">BBa_K1033910</a> - fwYellow, yellow chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033911">BBa_K1033911</a> - scOrange, orange chromoprotein (incl RBS, J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033912">BBa_K1033912</a> - scOrange, orange chromoprotein (incl RBS, J23106) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033913">BBa_K1033913</a> - scOrange, orange chromoprotein (incl. RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033914">BBa_K1033914</a> - amajLime, yellow-green chromoprotein (incl RBS & J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033915">BBa_K1033915</a> - amajLime, yellow-green chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033916">BBa_K1033916</a> - amajLime, yellow-green chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033917">BBa_K1033917</a> - gfasPurple, purple chromoprotein (incl RBS, J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033918">BBa_K1033918</a> - gfasPurple, purple chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033919">BBa_K1033919</a> - gfasPurple, purple chromoprotein <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033920">BBa_K1033920</a> - meffRed, red chromoprotein (incl RBS, J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033921">BBa_K1033921</a> - meffRed, red chromoprotein (incl RBS, J23106) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033922">BBa_K1033922</a> - meffRed, red chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033923">BBa_K1033923</a> - spisPink, pink chromoprotein (incl RBS, J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033924">BBa_K1033924</a> - spisPink, pink chromoprotein (incl RBS, J23106) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033925">BBa_K1033925</a> - spisPink, pink chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033926">BBa_K1033926</a> - asPink, pink chromoprotein (incl RBS, J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033927">BBa_K1033927</a> - asPink, pink chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033928">BBa_K1033928</a> - aeBlue, blue chromoprotein (incl RBS, J23110) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033929">BBa_K1033929</a> - aeBlue, blue chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033930">BBa_K1033930</a> - amilCP, blue/purple chromoprotein (incl RBS) <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033931">BBa_K1033931</a> - amilGFP, yellow chromoprotein (incl RBS) <br> <br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:UppsalaTeam:Uppsala2013-10-03T20:15:37Z<p>Sabrijamal: </p>
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<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
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<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/advisors">Advisors</a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/protocols">Protocols</a></li><br />
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Making the world healthier with a new probiotic platform<br />
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<p class="project-text"> Malnutrition is today a major global problem that affects people both in affluent and developing countries. Even if you get the right amount of calories, if these do not contain sufficient amounts of micronutrients, like vitamins and minerals, serious illness and even death can be the result.<br />
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The goal of our project is to alleviate this problem by applying synthetic biology to <a href="https://2013.igem.org/Team:Uppsala/probiotics">probiotic bacteria</a>. These bacteria are already a part of normal human diet and our goal is to modify them so that they enrich the nutritional content of the food they grow in.<br><br><br />
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With our project, we will make the lactobacillus genus the <a href="https://2013.igem.org/Team:Uppsala/chassi"> new probiotic platform </a> for <a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering"> metabolic engineering </a> of nutritional compounds. We will engineer probiotics to produce beta-carotene (provitamin), resveratrol (antioxidant), miraculin (sweetener) and saffron (taste).<br />
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To exemplify what this combination of probiotics and metabolic engineering can accomplish we hope to use our modified bacteria to create nutritionally enriched yoghurt. We have also put great effort into addressing the <a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics"> ethical and safety </a> issues that naturally follow when creating genetically modified food. As a part of this we organised a large event, <a href="https://2013.igem.org/Team:Uppsala/synbioday"> Uppsala SynBioDay </a>, with over 200 participants where these issues were discussed.<br />
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<h1>Sponsors</h1><br />
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<td><img class="sponsors" src="https://static.igem.org/mediawiki/2013/5/55/Uppsala_TekNat.png"></td><td><img class="sponsors" src="https://static.igem.org/mediawiki/2013/1/14/Uppsala_Analytik_Jena.png"></td></tr><br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:UppsalaTeam:Uppsala2013-10-03T20:13:04Z<p>Sabrijamal: </p>
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/advisors">Advisors</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/protocols">Protocols</a></li><br />
</ul></li><br />
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<div id="main_content"> <br />
<br />
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<div id="project-description"><br />
<h1><br />
Making the world healthier with a new probiotic platform<br />
</h1><br />
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<div id="text-box"><br />
<p class="project-text"> Malnutrition is today a major global problem that affects people both in affluent and developing countries. Even if you get the right amount of calories, if these do not contain sufficient amounts of micronutrients, like vitamins and minerals, serious illness and even death can be the result.<br />
<br />
The goal of our project is to alleviate this problem by applying synthetic biology to <a href="https://2013.igem.org/Team:Uppsala/probiotics">probiotic bacteria</a>. These bacteria are already a part of normal human diet and our goal is to modify them so that they enrich the nutritional content of the food they grow in.<br><br><br />
<br />
With our project, we will make the lactobacillus genus the <a href="https://2013.igem.org/Team:Uppsala/chassi"> new probiotic platform </a> for <a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering"> metabolic engineering </a> of nutritional compounds. We will engineer probiotics to produce beta-carotene (provitamin), resveratrol (antioxidant), miraculin (sweetener) and saffron (taste).<br />
<br />
To exemplify what this combination of probiotics and metabolic engineering can accomplish we hope to use our modified bacteria to create nutritionally enriched yoghurt. We have also put great effort into addressing the <a href="https://2013.igem.org/Team:Uppsala/safety">ethical and safety</a> issues that naturally follow when creating genetically modified food. As a part of this we organised a large event, <a href="https://2013.igem.org/Team:Uppsala/synbioday"> Uppsala SynBioDay </a>, with over 200 participants where these issues were discussed.<br />
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<div id="grid-a"><br />
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<a href="https://2013.igem.org/Team:Uppsala/project#project-anchor"><img class="top" src="https://static.igem.org/mediawiki/2013/a/a3/Uppsala2013_Background_new.png"></a> <br />
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<a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering"><img class="top" src="https://static.igem.org/mediawiki/2013/4/4f/Uppsala2013_Metabolicengineering.png"></a><br />
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<a href="https://2013.igem.org/Team:Uppsala/chassi"><img class="top" src="https://static.igem.org/mediawiki/2013/1/19/Uppsala2013_Chassi.png"></a><br />
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<a href="https://2013.igem.org/Team:Uppsala/human-practice"><img class="top" src="https://static.igem.org/mediawiki/2013/d/de/Uppsala2013_HumanPractice.png"></a><br />
</div><br />
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<a href="https://2013.igem.org/Team:Uppsala/results"><img class="top" src="https://static.igem.org/mediawiki/2013/a/a3/Uppsala2013_Achievements.png"></a><br />
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<h1>Sponsors</h1><br />
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<td class="centerThis"><img class="sponsors" src="https://static.igem.org/mediawiki/2013/a/a4/Uppsala_Clc.png"></td><br />
<td><img class="sponsors" src="https://static.igem.org/mediawiki/2013/5/55/Uppsala_TekNat.png"></td><td><img class="sponsors" src="https://static.igem.org/mediawiki/2013/1/14/Uppsala_Analytik_Jena.png"></td></tr><br />
<tr><td class="centerThis"><img class="sponsors" src="https://static.igem.org/mediawiki/2013/6/69/Uppsala_UU_logo.png"></td><td><img class="sponsors" src="https://static.igem.org/mediawiki/2013/9/9a/Uppsala_Microsynth.png"></td><td><img class="sponsors" src="https://static.igem.org/mediawiki/2013/b/ba/Vinnova.png"></td><br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:UppsalaTeam:Uppsala2013-10-03T20:12:28Z<p>Sabrijamal: </p>
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<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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</ul></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/advisors">Advisors</a></li><br />
<br />
</ul></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/protocols">Protocols</a></li><br />
</ul></li><br />
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<br />
<div id="main_content"> <br />
<br />
<br />
<br />
<div id="project-description"><br />
<h1><br />
Making the world healthier with a new probiotic platform<br />
</h1><br />
<br />
<div id="text-box"><br />
<p class="project-text"> Malnutrition is today a major global problem that affects people both in affluent and developing countries. Even if you get the right amount of calories, if these do not contain sufficient amounts of micronutrients, like vitamins and minerals, serious illness and even death can be the result.<br />
<br />
The goal of our project is to alleviate this problem by applying synthetic biology to <a href="https://2013.igem.org/Team:Uppsala/probiotics">probiotic bacteria</a>. These bacteria are already a part of normal human diet and our goal is to modify them so that they enrich the nutritional content of the food they grow in.<br><br><br />
<br />
With our project, we will make the lactobacillus genus the <a href="https://2013.igem.org/Team:Uppsala/chassi"> new probiotic platform </a> for <a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering"> metabolic engineering </a> of nutritional compounds. We will engineer probiotics to produce beta-carotene (provitamin), resveratrol (antioxidant), miraculin (sweetener) and saffron (taste).<br />
<br />
To exemplify what this combination of probiotics and metabolic engineering can accomplish we hope to use our modified bacteria to create nutritionally enriched <a href="https://2013.igem.org/Team:Uppsala/Yoghurtproducts">yoghurt</a>. We have also put great effort into addressing the <a href="https://2013.igem.org/Team:Uppsala/safety">ethical and safety</a> issues that naturally follow when creating genetically modified food. As a part of this we organised a large event, <a href="https://2013.igem.org/Team:Uppsala/synbioday"> Uppsala SynBioDay </a>, with over 200 participants where these issues were discussed.<br />
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<h1>Sponsors</h1><br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:UppsalaTeam:Uppsala2013-10-03T20:08:57Z<p>Sabrijamal: </p>
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<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/advisors">Advisors</a></li><br />
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</ul></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/protocols">Protocols</a></li><br />
</ul></li><br />
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<div id="main_content"> <br />
<br />
<br />
<br />
<div id="project-description"><br />
<h1><br />
Making the world healthier with a new probiotic platform<br />
</h1><br />
<br />
<div id="text-box"><br />
<p class="project-text"> Malnutrition is today a major global problem that affects people both in affluent and developing countries. Even if you get the right amount of calories, if these do not contain sufficient amounts of micronutrients, like vitamins and minerals, serious illness and even death can be the result.<br />
<br />
The goal of our project is to alleviate this problem by applying synthetic biology to <a href="https://2013.igem.org/Team:Uppsala/probiotics">probiotic bacteria</a>. These bacteria are already a part of normal human diet and our goal is to modify them so that they enrich the nutritional content of the food they grow in.<br><br><br />
<br />
With our project, we will make the <a href="https://2013.igem.org/Team:Uppsala/probiotics">lactobacillus genus</a> the new probiotic platform for metabolic engineering of nutritional compounds. We will engineer probiotics to produce <a href="https://2013.igem.org/Team:Uppsala/betacarotene">beta-carotene</a> (provitamin), <a href="https://2013.igem.org/Team:Uppsala/resveratrol">resveratrol</a> (antioxidant), <a href="https://2013.igem.org/Team:Uppsala/miraculin">miraculin</a> (sweetener) and <a href="https://2013.igem.org/Team:Uppsala/saffron">saffron</a> (taste).<br />
<br />
To exemplify what this combination of probiotics and metabolic engineering can accomplish we hope to use our modified bacteria to create nutritionally enriched <a href="https://2013.igem.org/Team:Uppsala/Yoghurtproducts">yoghurt</a>. We have also put great effort into addressing the <a href="https://2013.igem.org/Team:Uppsala/safety">ethical and safety</a> issues that naturally follow when creating genetically modified food. As a part of this we organised a large event, <a href="https://2013.igem.org/Team:Uppsala/synbioday"> Uppsala SynBioDay </a>, with over 200 participants where these issues were discussed.<br />
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<h1>Sponsors</h1><br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/vectorsTeam:Uppsala/vectors2013-10-03T20:06:32Z<p>Sabrijamal: </p>
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<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
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<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title">Shuttle vectors for Lactobacillus and E. coli</h1><br />
<div id="p-com-text1"><br />
<h1>Building bridges between E. coli and Lactobacillus</h1><br />
<br />
<p>A shuttle vector is a plasmid that can be transferred between two different species and is able to replicate in both. Such a vector is key for bioengineering in Lactobacillus. The low transformation frequencies of ligations and longer generation times slows down the work pace tremendously if you have to assembly and test everything in Lactobacillus. A solution to this problem is if you instead build everything in E.coli and then transfer the finished construct to Lactobacillus. We have created two shuttle vectors that we have sent to the registry.<br><br><br />
<br />
Resveratrol belongs to a group of molecules known as Phyloalexin which is used by many plants to battle infections of all sorts.<br />
Since the early 90’s there has been many studies done on resveratrol showing it has a wide range of beneficial properties ranging from skin cancer reduction to anti-inflamatory effects and antioxidant properties.<br><br><br />
<br />
<h3>Shuttle vectors</h3><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206, with chloramphenicol resistance.</a><br><br />
<a href="http://parts.igem.org/Part:BBa_K1033207">BBa_K1033207, with erythromycin resistance.</a><br />
<br><br><br />
<br />
</div><br />
<div class="text"><br />
<a id="b1"></a><h1>Construction of shuttle vectorns</h1><br />
<p><br />
To create the shuttle vector we replaced the replicon in a biobrick compatible plasmid BBa_K864003?) with a replicon from a plasmid known to work in both E.coli and Lactobacillus pSH71. pSH71 originates from a plasmid, pJP059, from Lactococcus lactis but it is well known to replicate in both the Lactobacillus genus and in E.coli through rolling circle replication.<sup><a href="#ref_point">[1]</a></sup><br><br><br />
<br />
Many species in the Lactobacillus genus have inherent antibiotic resistances.<sup><a href="#ref_point">[2]</a></sup>, because of that we have been limited in our selection of antibiotic resistance. Most commonly used are chloramphenicol and erythromycin and we made versions of both for our shuttle vector. BBa_K1033207 contains an erythromycin cassette from pLUL631 and BBa_K1033206 contains the resistance cassette from pSB1C3 but has had its promoter replaced with our cp29 promoter.<sup><a href="#ref_point">[3]</a></sup><br><br> </p><br />
</div><br />
<img class="vector-plasmid" src="https://static.igem.org/mediawiki/2013/9/92/Shuttle-vector_pSBLbC.png"><br />
<br />
<h1>Results</h1><br />
<br />
<p>We have managed to transform both shuttle vectors to E. coli and verified them by sequencing. We have also managed to subclone with them and replaced the red insert with chromoproteins expressed by our CP promoters that works both in E. coli and Lactobacillus, clearly yielding coloured colonies (see picture on the right below).</p><br />
<br />
<img class="pic_type2" src="https://static.igem.org/mediawiki/2013/e/e1/IMG_6644.JPG"><br />
<img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"><br />
<br />
<p>Finally we have also gained positive results when transforming BBa_K1033207 to Lactobacillus reuteri and Lactobacillus plantarum. See the picture above on the left. The plate on the left contains Lactobacillus reuteri 100-23 transformed with BBa_K1033207. The plate on the right is a negative control of 100-23 without the plasmid. <p><br />
<br />
<div id="reference"><br />
<h1>Reference</h1><br />
<p class="reference"><br />
<br />
<a>[1]</a> <a href:"http://www.ncbi.nlm.nih.gov/pubmed/11591136> "Construction of compatible wide-host-range shuttle v... </a> [Plasmid. 2001] - PubMed - NCBI." National Center for Biotechnology Information. N.p., n.d. Web. 1 Oct. 2013. <br />
<br />
<br><br><br />
<br />
<a>[2]</a> <a href:"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3378904> "High efficiency recombineering in lactic acid bacteria." </a> National Center for Biotechnology Information. N.p., n.d. Web. 1 Oct. 2013.<br />
<br><br><br />
<br />
<a>[3]</a> <a href="Transformation of Lactobacillus reuteri with electroporation: Studies on the erythromycin resistance plasmid pLUL631 , Siv Ahrné, Göran Molin, Lars Axelsson, Januari 1992"> Transformation of Lactobacillus reuteri with electroporation: Studies on the erythromycin resistance plasmid pLUL631 </a> , Siv Ahrné, Göran Molin, Lars Axelsson, Januari 1992<br />
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}</div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/vectorsTeam:Uppsala/vectors2013-10-03T20:01:02Z<p>Sabrijamal: </p>
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<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
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<li><a <br />
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<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
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<h1 class="main-title">Shuttle vectors for Lactobacillus and E. coli</h1><br />
<div id="p-com-text1"><br />
<h1>Building bridges between E. coli and Lactobacillus</h1><br />
<br />
<p>A shuttle vector is a plasmid that can be transferred between two different species and is able to replicate in both. Such a vector is key for bioengineering in Lactobacillus. The low transformation frequencies of ligations and longer generation times slows down the work pace tremendously if you have to assembly and test everything in Lactobacillus. A solution to this problem is if you instead build everything in E.coli and then transfer the finished construct to Lactobacillus. We have created two shuttle vectors that we have sent to the registry.<br><br><br />
<br />
Resveratrol belongs to a group of molecules known as Phyloalexin which is used by many plants to battle infections of all sorts.<br />
Since the early 90’s there has been many studies done on resveratrol showing it has a wide range of beneficial properties ranging from skin cancer reduction to anti-inflamatory effects and antioxidant properties.<br><br><br />
<br />
<h3>Shuttle vectors</h3><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206, with chloramphenicol resistance.</a><br><br />
<a href="http://parts.igem.org/Part:BBa_K1033207">BBa_K1033207, with erythromycin resistance.</a><br />
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<div class="text"><br />
<a id="b1"></a><h1>Construction of shuttle vectorns</h1><br />
<p><br />
To create the shuttle vector we replaced the replicon in a biobrick compatible plasmid BBa_K864003?) with a replicon from a plasmid known to work in both E.coli and Lactobacillus pSH71. pSH71 originates from a plasmid, pJP059, from Lactococcus lactis but it is well known to replicate in both the Lactobacillus genus and in E.coli through rolling circle replication.<sup><a href="#ref_point">[1]</a></sup><br><br><br />
<br />
Many species in the Lactobacillus genus have inherent antibiotic resistances.<sup><a href="#ref_point">[2]</a></sup>, because of that we have been limited in our selection of antibiotic resistance. Most commonly used are chloramphenicol and erythromycin and we made versions of both for our shuttle vector. BBa_K1033207 contains an erythromycin cassette from pLUL631 and BBa_K1033206 contains the resistance cassette from pSB1C3 but has had its promoter replaced with our cp29 promoter.<sup><a href="#ref_point">[3]</a></sup><br><br> </p><br />
</div><br />
<img class="pic_type1" src="https://static.igem.org/mediawiki/2013/9/92/Shuttle-vector_pSBLbC.png"><br />
<br />
<h1>Results</h1><br />
<br />
<p>We have managed to transform both shuttle vectors to E. coli and verified them by sequencing. We have also managed to subclone with them and replaced the red insert with chromoproteins expressed by our CP promoters that works both in E. coli and Lactobacillus, clearly yielding coloured colonies (see picture on the right below).</p><br />
<br />
<table border="1"><br />
<tr><br />
<td> <img class="pic_type2" src="https://static.igem.org/mediawiki/2013/e/e1/IMG_6644.JPG"> </td><br />
<td> <img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </td><br />
</tr><br />
</table><br />
<br />
<p>Finally we have also gained positive results when transforming BBa_K1033207 to Lactobacillus reuteri and Lactobacillus plantarum. See the picture above on the left. The plate on the left contains Lactobacillus reuteri 100-23 transformed with BBa_K1033207. The plate on the right is a negative control of 100-23 without the plasmid. <p><br />
<br />
<div id="reference"><br />
<h1>Reference</h1><br />
<p class="reference"><br />
<br />
<a>[1]</a> <a href:"http://www.ncbi.nlm.nih.gov/pubmed/11591136> "Construction of compatible wide-host-range shuttle v... </a> [Plasmid. 2001] - PubMed - NCBI." National Center for Biotechnology Information. N.p., n.d. Web. 1 Oct. 2013. <br />
<br />
<br><br><br />
<br />
<a>[2]</a> <a href:"http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3378904> "High efficiency recombineering in lactic acid bacteria." </a> National Center for Biotechnology Information. N.p., n.d. Web. 1 Oct. 2013.<br />
<br><br><br />
<br />
<a>[3]</a> <a href="Transformation of Lactobacillus reuteri with electroporation: Studies on the erythromycin resistance plasmid pLUL631 , Siv Ahrné, Göran Molin, Lars Axelsson, Januari 1992"> Transformation of Lactobacillus reuteri with electroporation: Studies on the erythromycin resistance plasmid pLUL631 </a> , Siv Ahrné, Göran Molin, Lars Axelsson, Januari 1992<br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T20:00:04Z<p>Sabrijamal: </p>
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
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<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus. Below in the picture is a plate with our shuttle vector <a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206</a> containing the chromoprotein amilCP fused with the synthetic promoter CP29 from our promoter collection. The colonies are clearly blue efter 24 hours in E. coli, we have not yet been able to transform the construct to Lactobacillus.</p><br />
<br><br />
<br />
<img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> <br />
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<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/promotersTeam:Uppsala/promoters2013-10-03T19:53:03Z<p>Sabrijamal: </p>
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<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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</ul></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
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</ul></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
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<br />
<h1 class="main-title"> Constitutive promoter library for Lactobacillus & E.coli </h1><br />
<br />
<h1> Introducing new constitutive promoters to the iGEM community </h1><br />
<br />
Depending on how much you want to express your gene, you will need promoters of different strengths. Therefore we wanted to provide a selection of promoters that work in both E. coli and Lactobacillus. We synthesized seven promoters based on the CP promoter library in the article<sup><a href="#ref_point">[1]</a></sup>. These promoters are based on consensus sequences from promoters in Lactococcus lactis. Below you can see our promoters and their name in the part registry. <br />
<br />
<br><br />
<br><br />
<br />
<li> CP1 - <a href="http://parts.igem.org/Part:BBa_K1033219"> BBa_K1033219 </a> </li><br />
<li> CP8 - <a href="http://parts.igem.org/Part:BBa_K1033220"> BBa_K1033220 </a> </li><br />
<li> CP11 - <a href="http://parts.igem.org/Part:BBa_K1033221"> BBa_K1033221 </a> </li><br />
<li> CP29 - <a href="http://parts.igem.org/Part:BBa_K1033222"> BBa_K1033222 </a> </li><br />
<li> CP30 - <a href="http://parts.igem.org/Part:BBa_K1033223"> BBa_K1033223 </a> </li><br />
<li> CP41 - <a href="http://parts.igem.org/Part:BBa_K1033224"> BBa_K1033224 </a> </li><br />
<li> CP44 - <a href="http://parts.igem.org/Part:BBa_K1033225">BBa_K1033225 </a> </li><br />
<br />
<br> <br />
<br />
<a id="b1"></a><br />
<div id="promoter_fac"> <img class="promoter_image" src="https://static.igem.org/mediawiki/2013/7/72/Uppsala2013_Facspic.png" > <br />
<div id="promoter_fb"> <br> <i> Strength measuring of CP1 and CP41 in E. coli using FACS. – The peaks show the median of how much the <br><br />
cells in the sample fluoresced. </i> </div<br />
</div><br />
<br />
<br />
<br><br />
<br><br />
<br />
To characterize them we assembled them with the reporter gene <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K592024"> BBa_K592024 </a> B0034-BFP in pSB3K3 and transformed them into E. coli. BFP stands for “Blue Fluorescence Protein” and the expression of this gene leads to production of a fluorescence protein which we can measure with a fluorescence-activated cell sorting (FACS) machine. The FACS gives you a value of the fluorescence from each cell in your sample. As a reference we used the promoter J23101 from the standard parts in the registry, where its strength were put to 1.<br />
<br />
<br><br />
<br><br />
<br />
<div id="promoter_bar"> <a id="l1"><img class="promoter_image" src="https://static.igem.org/mediawiki/2013/6/64/Uppsala2013_Promoterdiagram.png"></a> <br />
<div id="promoter_fb"> <br> <i> The CP promoters strength relative to J23101 [RPU] in E. coli. </i> </div><br />
</div> <br />
<br />
<br />
<br />
<br><br />
<br><br />
From this we got the relative strength of our promoters, compared to J23101. The weakest promoters in our collection have half the strength of J23101, while the strongest promoter is approximately four times stronger. <br />
<br />
<br><br />
<br><br />
<br />
<h1> Further characterization of CP promoters in Lactobacillus</h1><br />
<br />
Because of time constraints, we have not had the time to characterize them in Lactobacillus. Though in the future we plan to do this by first sub cloning them together with <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K592024"> BBa_K592024 </a>B0034-BFP from our previous assemblies and then assemble them with our shuttle vector. After that we can transform them into either Lactobacillus reuteri or Lactobacillus plantarum and characterize them in the same way as we did in E. coli. The only difference will be that we cant’t be sure if J23101 will work in L. reuteri or L. plantarum. In that case we will only get the strength of each promoter relative the strength of our other CP promoters.<br />
<br> <br />
<br><br />
<br />
These promoters have been used successfully in several parts in our project, including <a href="http://parts.igem.org/Part:BBa_K1033000">BBa_K1033000</a> (TAL), <a href="http://parts.igem.org/Part:BBa_K1033002">BBa_K1033002</a> (STS) and <a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> amilCP.<br />
<br />
<br> <br />
<br><br />
<br />
In the article by Jensen and Hammer they stated that these promoters were based on the consensus sequence of promoters in gram-positive bacterium L. lactis, though there are no obvious reasons why they shouldn’t work in other organisms. Jensen and Hammer characterized them in both L. lactis and gram-negative bacterium E. coli using the reporter gene lacLM. This indicates that there is a possibility that these promoters are near universally applicable to prokaryotic organisms in general.<br />
<a name="ref_point"></a><br />
<h1>References</h1><br />
<a>[1].</a> The Sequence of Spacers between the Consensus Sequences<br />
Modulates the Strength of Prokaryotic Promoters<br />
PETER RUHDAL JENSEN AND KARIN HAMMER<br />
Department of Microbiology, Technical University of Denmark, DK-2800 Lyngby, Denmark 21 October 1997 - <a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC124675/pdf/am000082.pdf">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC124675/pdf/am000082.pdf</a><br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/promotersTeam:Uppsala/promoters2013-10-03T19:46:46Z<p>Sabrijamal: </p>
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title"> Constitutive promoter library for Lactobacillus & E.coli </h1><br />
<br />
<h1> Introducing new constitutive promoters to the iGEM community </h1><br />
<br />
Depending on how much you want to express your gene, you will need promoters of different strengths. Therefore we wanted to provide a selection of promoters that work in both E. coli and Lactobacillus. We synthesized seven promoters based on the CP promoter library in the article<sup><a href="#ref_point">[1]</a></sup>. These promoters are based on consensus sequences from promoters in Lactococcus lactis. Below you can see our promoters and their name in the part registry. <br />
<br />
<br><br />
<br><br />
<br />
<li> CP1 - <a href="http://parts.igem.org/Part:BBa_K1033219"> BBa_K1033219 </a> </li><br />
<li> CP8 - <a href="http://parts.igem.org/Part:BBa_K1033220"> BBa_K1033220 </a> </li><br />
<li> CP11 - <a href="http://parts.igem.org/Part:BBa_K1033221"> BBa_K1033221 </a> </li><br />
<li> CP29 - <a href="http://parts.igem.org/Part:BBa_K1033222"> BBa_K1033222 </a> </li><br />
<li> CP30 - <a href="http://parts.igem.org/Part:BBa_K1033223"> BBa_K1033223 </a> </li><br />
<li> CP41 - <a href="http://parts.igem.org/Part:BBa_K1033224"> BBa_K1033224 </a> </li><br />
<li> CP44 - <a href="http://parts.igem.org/Part:BBa_K1033225">BBa_K1033225 </a> </li><br />
<br />
<br> <br />
<br />
<a id="b1"></a><br />
<div id="promoter_fac"> <img class="promoter_image" src="https://static.igem.org/mediawiki/2013/7/72/Uppsala2013_Facspic.png" > <br />
<div id="promoter_fb"> <br> <i> Strength measuring of CP1 and CP41 in E. coli using FACS. – The peaks show the median of how much the <br><br />
cells in the sample fluoresced. </i> </div<br />
</div><br />
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<br />
<br><br />
<br><br />
<br />
To characterize them we assembled them with the reporter gene <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K592024"> BBa_K592024 </a> B0034-BFP in pSB3K3 and transformed them into E. coli. BFP stands for “Blue Fluorescence Protein” and the expression of this gene leads to production of a fluorescence protein which we can measure with a fluorescence-activated cell sorting (FACS) machine. The FACS gives you a value of the fluorescence from each cell in your sample. As a reference we used the promoter J23101 from the standard parts in the registry, where its strength were put to 1.<br />
<br />
<br><br />
<br><br />
<br />
<div id="promoter_bar"> <a id="l1"><img class="promoter_image" src="https://static.igem.org/mediawiki/2013/6/64/Uppsala2013_Promoterdiagram.png"></a> <br />
<div id="promoter_fb"> <br> <i> The CP promoters strength relative to J23101 [RPU] in E. coli. </i> </div><br />
</div> <br />
<br />
<br />
<br />
<br><br />
<br><br />
From this we got the relative strength of our promoters, compared to J23101. The weakest promoters in our collection have half the strength of J23101, while the strongest promoter is approximately four times stronger. <br />
<br />
<br><br />
<br><br />
<br />
<h1> Further characterization of CP promoters in Lactobacillus</h1><br />
<br />
Because of time constraints, we have not had the time to characterize them in Lactobacillus. Though in the future we plan to do this by first sub cloning them together with <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K592024"> BBa_K592024 </a>B0034-BFP from our previous assemblies and then assemble them with our shuttle vector. After that we can transform them into either Lactobacillus reuteri or Lactobacillus plantarum and characterize them in the same way as we did in E. coli. The only difference will be that we cant’t be sure if J23101 will work in L. reuteri or L. plantarum. In that case we will only get the strength of each promoter relative the strength of our other CP promoters.<br />
<br />
These promoters have been used successfully in several parts in our project, including TAL, STS and amilCP.<br />
<br />
<br> <br />
<br><br />
<br />
In the article by Jensen and Hammer they stated that these promoters were based on the consensus sequence of promoters in gram-positive bacterium L. lactis, though there are no obvious reasons why they shouldn’t work in other organisms. Jensen and Hammer characterized them in both L. lactis and gram-negative bacterium E. coli using the reporter gene lacLM. This indicates that there is a possibility that these promoters are near universally applicable to prokaryotic organisms in general.<br />
<a name="ref_point"></a><br />
<h1>References</h1><br />
<a>[1].</a> The Sequence of Spacers between the Consensus Sequences<br />
Modulates the Strength of Prokaryotic Promoters<br />
PETER RUHDAL JENSEN AND KARIN HAMMER<br />
Department of Microbiology, Technical University of Denmark, DK-2800 Lyngby, Denmark 21 October 1997 - <a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC124675/pdf/am000082.pdf">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC124675/pdf/am000082.pdf</a><br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:42:36Z<p>Sabrijamal: </p>
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
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<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
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<h1 class="main-title-left">Reporter genes</h1><br />
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<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
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<div id="clear"></div><br />
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<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus. Below in the picture is a plate with our shuttle vector <a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206</a> containing the chromoprotein amilCP fused with the synthetic promoter CP29 from our promoter collection. The colonies are clearly blue efter 24 hours in E. coli, we have not yet been able to transform the construct to Lactobacillus.</p><br />
<br><br />
<br />
<img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img> <br />
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<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:40:49Z<p>Sabrijamal: </p>
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
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<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus. Our shuttle vector <a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206</a> containing the chromoprotein amilCP fused with the synthetic promoter CP29 from our promoter collection. The colonies are clearly blue efter 24 hours in E. coli, we have not yet been able to transform the construct to Lactobacillus.</p><br />
<br><br />
<br />
<img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img> <br />
<br />
<br />
<!--<br />
<div id="shuttle_vec_cont"> <br />
<br />
</div><br />
--><br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
<br />
</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
<br />
</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:34:23Z<p>Sabrijamal: </p>
<hr />
<div>{{Team:Uppsala/repgenes-style}}<br />
{{Team:Uppsala/standard-style-no-frame}}<br />
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<div id="igem_logo"><a href="https://2013.igem.org/Team:Uppsala"><img class="igem_logo" src="https://static.igem.org/mediawiki/2013/e/e0/Uppsala2013_IGEMlogo.png"></a></div> <br />
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<div id="igem"><a href="https://2013.igem.org/Main_Page"><img class="igem" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_Uppsala_IGEM_log_blue.png"></a></div><br />
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<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus. Our shuttle vector <a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206</a> containing the chromoprotein amilCP fused with the synthetic promoter CP29 from our promoter collection. The colonies are clearly blue efter 24 hours in E. coli, we have not yet been able to transform the construct to Lactobacillus.</p><br />
<br />
<br><br />
<!--<br />
<table border="0"><br />
<tr><br />
<td> <img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img> </td><br />
</tr><br />
<tr> <br />
<td> <i> </i> </td><br />
</tr><br />
</table><br />
--><br />
<br />
<!--<br />
<div id="shuttle_vec_cont"> <br />
<br />
</div><br />
--><br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
<br />
</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
<br />
</html></div>Sabrijamalhttp://2013.igem.org/Template:Team:Uppsala/repgenes-styleTemplate:Team:Uppsala/repgenes-style2013-10-03T19:31:23Z<p>Sabrijamal: </p>
<hr />
<div><html><br />
<style type="text/css"><br />
<br />
#main_frame {<br />
min-height: 1300px;<br />
width: 950px;<br />
margin: 20px auto;<br />
border: 1px solid ligth gray;<br />
border-radius: 15px;<br />
box-shadow: 0px 0px 30px rgba(110,110,110, .8); <br />
background: white;<br />
<br />
}<br />
<br />
.main-title {<br />
<br />
border:none;<br />
font-size:38px;<br />
color:#22409a;<br />
<br />
}<br />
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.main-title-left {<br />
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border:none;<br />
font-size:38px;<br />
color:#22409a;<br />
float:left;<br />
<br />
}<br />
<br />
.second-row {<br />
border:none;<br />
font-size:38px;<br />
color:#22409a;<br />
position:relative;<br />
top:-20px;<br />
}<br />
<br />
.no-border {<br />
border:none;<br />
}<br />
<br />
h1 {<br />
color:#0089d1;<br />
}<br />
<br />
h3 {<br />
color:#0089d1;<br />
}<br />
<br />
.symbol-pic {<br />
float:right;<br />
width:50px;<br />
height:50px;<br />
}<br />
<br />
#clear {<br />
clear:both;<br />
}<br />
<br />
.shuttle_vec{<br />
width:362px;<br />
height:272px;<br />
margin-left:300px;<br />
}<br />
<br />
#shuttle_vec_cont{<br />
width:362px;<br />
height:390px;<br />
margin-left:300px;<br />
}<br />
<br />
</style></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:30:38Z<p>Sabrijamal: </p>
<hr />
<div>{{Team:Uppsala/repgenes-style}}<br />
{{Team:Uppsala/standard-style-no-frame}}<br />
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<br />
<html><br />
<br />
<head><br />
<meta charset="utf-8" /> <br />
<title>Uppsala iGEM 2013</title> <br />
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<br />
<div id= "main_frame"> <br />
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<!--Contains our top header--><br />
<br />
<div id= "header"> <br />
<br />
<!-- Contains sub-logo picture --><br />
<div id="igem_logo"><a href="https://2013.igem.org/Team:Uppsala"><img class="igem_logo" src="https://static.igem.org/mediawiki/2013/e/e0/Uppsala2013_IGEMlogo.png"></a></div> <br />
<br />
<div id="igem"><a href="https://2013.igem.org/Main_Page"><img class="igem" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_Uppsala_IGEM_log_blue.png"></a></div><br />
<br />
<img class="slogan" src="https://static.igem.org/mediawiki/2013/a/ad/Uppsala2013_Header.png"> <br />
<br />
</div> <br />
<br />
<!-- Contains the "nav div" and the stripe background for the navigation menu --> <br />
<br />
<div id="navbar"> <br />
<br />
<!-- Contains the navigation menu with list elements, buged on local server, works on mediawiki!!--> <br />
<br />
<div id="nav"><br />
<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<br><br />
<br />
<table border="0"><br />
<tr><br />
<td> <img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img> </td><br />
</tr><br />
<tr> <br />
<td> <i> Our shuttle vector <a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206</a> containing the chromoprotein amilCP fused with the synthetic promoter CP29 from our promoter collection. The colonies are clearly blue efter 24 hours in E. coli, we have not yet been able to transform the construct to Lactobacillus. </i> </td><br />
</tr><br />
</table><br />
<br />
<!--<br />
<div id="shuttle_vec_cont"> <br />
<br />
</div><br />
--><br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
<br />
</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
<br />
</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:29:19Z<p>Sabrijamal: </p>
<hr />
<div>{{Team:Uppsala/repgenes-style}}<br />
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<br />
<div id="igem"><a href="https://2013.igem.org/Main_Page"><img class="igem" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_Uppsala_IGEM_log_blue.png"></a></div><br />
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<br />
<div id="navbar"> <br />
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<!-- Contains the navigation menu with list elements, buged on local server, works on mediawiki!!--> <br />
<br />
<div id="nav"><br />
<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<br><br />
<br />
<table border="0"><br />
<tr><br />
<td> <img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img> </td><br />
<td><br> <br> <br> <br> <br> <br> <br> <br> <br> <br> <br> <br> <i> Our shuttle vector <a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206</a> containing the chromoprotein amilCP fused with the synthetic promoter CP29 from our promoter collection. The colonies are clearly blue efter 24 hours in E. coli, we have not yet been able to transform the construct to Lactobacillus. </i> </td><br />
</tr><br />
</table><br />
<br />
<!--<br />
<div id="shuttle_vec_cont"> <br />
<br />
</div><br />
--><br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
<br />
</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
<br />
</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:29:02Z<p>Sabrijamal: </p>
<hr />
<div>{{Team:Uppsala/repgenes-style}}<br />
{{Team:Uppsala/standard-style-no-frame}}<br />
<br />
<br />
<html><br />
<br />
<head><br />
<meta charset="utf-8" /> <br />
<title>Uppsala iGEM 2013</title> <br />
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<br />
<div id= "main_frame"> <br />
<br />
<!--Contains our top header--><br />
<br />
<div id= "header"> <br />
<br />
<!-- Contains sub-logo picture --><br />
<div id="igem_logo"><a href="https://2013.igem.org/Team:Uppsala"><img class="igem_logo" src="https://static.igem.org/mediawiki/2013/e/e0/Uppsala2013_IGEMlogo.png"></a></div> <br />
<br />
<div id="igem"><a href="https://2013.igem.org/Main_Page"><img class="igem" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_Uppsala_IGEM_log_blue.png"></a></div><br />
<br />
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<br />
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<br />
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<br />
<div id="navbar"> <br />
<br />
<!-- Contains the navigation menu with list elements, buged on local server, works on mediawiki!!--> <br />
<br />
<div id="nav"><br />
<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<br><br />
<br />
<table border="0"><br />
<tr><br />
<td> <img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img> </td><br />
<td><br> <br> <br> <br> <br> <br> <br> <br> <i> Our shuttle vector <a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206</a> containing the chromoprotein amilCP fused with the synthetic promoter CP29 from our promoter collection. The colonies are clearly blue efter 24 hours in E. coli, we have not yet been able to transform the construct to Lactobacillus. </i> </td><br />
</tr><br />
</table><br />
<br />
<!--<br />
<div id="shuttle_vec_cont"> <br />
<br />
</div><br />
--><br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
<br />
</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
<br />
</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:28:35Z<p>Sabrijamal: </p>
<hr />
<div>{{Team:Uppsala/repgenes-style}}<br />
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<br />
<div id="navbar"> <br />
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<!-- Contains the navigation menu with list elements, buged on local server, works on mediawiki!!--> <br />
<br />
<div id="nav"><br />
<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<br><br />
<br />
<table border="0"><br />
<tr><br />
<td> <img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img> </td><br />
<td><br> <br> <br> <br> <i> Our shuttle vector <a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206</a> containing the chromoprotein amilCP fused with the synthetic promoter CP29 from our promoter collection. The colonies are clearly blue efter 24 hours in E. coli, we have not yet been able to transform the construct to Lactobacillus. </i> </td><br />
</tr><br />
</table><br />
<br />
<!--<br />
<div id="shuttle_vec_cont"> <br />
<br />
</div><br />
--><br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
<br />
</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
<br />
</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:27:46Z<p>Sabrijamal: </p>
<hr />
<div>{{Team:Uppsala/repgenes-style}}<br />
{{Team:Uppsala/standard-style-no-frame}}<br />
<br />
<br />
<html><br />
<br />
<head><br />
<meta charset="utf-8" /> <br />
<title>Uppsala iGEM 2013</title> <br />
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<div id= "main_frame"> <br />
<br />
<!--Contains our top header--><br />
<br />
<div id= "header"> <br />
<br />
<!-- Contains sub-logo picture --><br />
<div id="igem_logo"><a href="https://2013.igem.org/Team:Uppsala"><img class="igem_logo" src="https://static.igem.org/mediawiki/2013/e/e0/Uppsala2013_IGEMlogo.png"></a></div> <br />
<br />
<div id="igem"><a href="https://2013.igem.org/Main_Page"><img class="igem" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_Uppsala_IGEM_log_blue.png"></a></div><br />
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<br />
<div id="navbar"> <br />
<br />
<!-- Contains the navigation menu with list elements, buged on local server, works on mediawiki!!--> <br />
<br />
<div id="nav"><br />
<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<br><br />
<br />
<table border="0"><br />
<tr><br />
<td> <img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img> </td><br />
<td> <i> Our shuttle vector <a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206</a> containing the chromoprotein amilCP fused with the synthetic promoter CP29 from our promoter collection. The colonies are clearly blue efter 24 hours in E. coli, we have not yet been able to transform the construct to Lactobacillus. </i> </td><br />
</tr><br />
</table><br />
<br />
<!--<br />
<div id="shuttle_vec_cont"> <br />
<br />
</div><br />
--><br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
<br />
</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
<br />
</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:26:09Z<p>Sabrijamal: </p>
<hr />
<div>{{Team:Uppsala/repgenes-style}}<br />
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<br />
<div id="igem"><a href="https://2013.igem.org/Main_Page"><img class="igem" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_Uppsala_IGEM_log_blue.png"></a></div><br />
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<br />
<div id="navbar"> <br />
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<!-- Contains the navigation menu with list elements, buged on local server, works on mediawiki!!--> <br />
<br />
<div id="nav"><br />
<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<br><br />
<br />
<table border="0"><br />
<tr><br />
<td> <img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img> </td><br />
</tr><br />
<tr><br />
<td> <i> Our shuttle vector <a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206</a> containing the chromoprotein amilCP fused with the synthetic promoter CP29 from our promoter collection. The colonies are clearly blue efter 24 hours in E. coli, we have not yet been able to transform the construct to Lactobacillus. </i> </td><br />
</tr><br />
</table><br />
<br />
<!--<br />
<div id="shuttle_vec_cont"> <br />
<br />
</div><br />
--><br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
<br />
</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
<br />
</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:25:47Z<p>Sabrijamal: </p>
<hr />
<div>{{Team:Uppsala/repgenes-style}}<br />
{{Team:Uppsala/standard-style-no-frame}}<br />
<br />
<br />
<html><br />
<br />
<head><br />
<meta charset="utf-8" /> <br />
<title>Uppsala iGEM 2013</title> <br />
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<br />
<div id= "main_frame"> <br />
<br />
<!--Contains our top header--><br />
<br />
<div id= "header"> <br />
<br />
<!-- Contains sub-logo picture --><br />
<div id="igem_logo"><a href="https://2013.igem.org/Team:Uppsala"><img class="igem_logo" src="https://static.igem.org/mediawiki/2013/e/e0/Uppsala2013_IGEMlogo.png"></a></div> <br />
<br />
<div id="igem"><a href="https://2013.igem.org/Main_Page"><img class="igem" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_Uppsala_IGEM_log_blue.png"></a></div><br />
<br />
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<!-- Contains the "nav div" and the stripe background for the navigation menu --> <br />
<br />
<div id="navbar"> <br />
<br />
<!-- Contains the navigation menu with list elements, buged on local server, works on mediawiki!!--> <br />
<br />
<div id="nav"><br />
<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<table border="0"><br />
<tr><br />
<td> <img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img> </td><br />
</tr><br />
<tr><br />
<td> <i> Our shuttle vector <a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206</a> containing the chromoprotein amilCP fused with the synthetic promoter CP29 from our promoter collection. The colonies are clearly blue efter 24 hours in E. coli, we have not yet been able to transform the construct to Lactobacillus. </i> </td><br />
</tr><br />
</table><br />
<br />
<!--<br />
<div id="shuttle_vec_cont"> <br />
<br />
</div><br />
--><br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
<br />
</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
<br />
</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:25:14Z<p>Sabrijamal: </p>
<hr />
<div>{{Team:Uppsala/repgenes-style}}<br />
{{Team:Uppsala/standard-style-no-frame}}<br />
<br />
<br />
<html><br />
<br />
<head><br />
<meta charset="utf-8" /> <br />
<title>Uppsala iGEM 2013</title> <br />
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<style type="text/css"><br />
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<!--Main frame contains all the div elements--><br />
<br />
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<br />
<!--Contains our top header--><br />
<br />
<div id= "header"> <br />
<br />
<!-- Contains sub-logo picture --><br />
<div id="igem_logo"><a href="https://2013.igem.org/Team:Uppsala"><img class="igem_logo" src="https://static.igem.org/mediawiki/2013/e/e0/Uppsala2013_IGEMlogo.png"></a></div> <br />
<br />
<div id="igem"><a href="https://2013.igem.org/Main_Page"><img class="igem" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_Uppsala_IGEM_log_blue.png"></a></div><br />
<br />
<img class="slogan" src="https://static.igem.org/mediawiki/2013/a/ad/Uppsala2013_Header.png"> <br />
<br />
</div> <br />
<br />
<!-- Contains the "nav div" and the stripe background for the navigation menu --> <br />
<br />
<div id="navbar"> <br />
<br />
<!-- Contains the navigation menu with list elements, buged on local server, works on mediawiki!!--> <br />
<br />
<div id="nav"><br />
<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<table border="1"><br />
<tr><br />
<td>row 1, cell 1</td><br />
<td>row 1, cell 2</td><br />
</tr><br />
<tr><br />
<td>row 2, cell 1</td><br />
<td>row 2, cell 2</td><br />
</tr><br />
</table><br />
<br />
<br />
<table border="1"><br />
<tr><br />
<td> <img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img> </td><br />
</tr><br />
<tr><br />
<td> <i> Our shuttle vector <a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206</a> containing the chromoprotein amilCP fused with the synthetic promoter CP29 from our promoter collection. The colonies are clearly blue efter 24 hours in E. coli, we have not yet been able to transform the construct to Lactobacillus. </i> </td><br />
</tr><br />
</table><br />
<br />
<!--<br />
<div id="shuttle_vec_cont"> <br />
<br />
</div><br />
--><br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
<br />
</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
<br />
</html></div>Sabrijamalhttp://2013.igem.org/Template:Team:Uppsala/repgenes-styleTemplate:Team:Uppsala/repgenes-style2013-10-03T19:17:33Z<p>Sabrijamal: </p>
<hr />
<div><html><br />
<style type="text/css"><br />
<br />
#main_frame {<br />
min-height: 1300px;<br />
width: 950px;<br />
margin: 20px auto;<br />
border: 1px solid ligth gray;<br />
border-radius: 15px;<br />
box-shadow: 0px 0px 30px rgba(110,110,110, .8); <br />
background: white;<br />
<br />
}<br />
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.main-title {<br />
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font-size:38px;<br />
color:#22409a;<br />
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}<br />
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border:none;<br />
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color:#22409a;<br />
float:left;<br />
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}<br />
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.second-row {<br />
border:none;<br />
font-size:38px;<br />
color:#22409a;<br />
position:relative;<br />
top:-20px;<br />
}<br />
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border:none;<br />
}<br />
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h1 {<br />
color:#0089d1;<br />
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h3 {<br />
color:#0089d1;<br />
}<br />
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.symbol-pic {<br />
float:right;<br />
width:50px;<br />
height:50px;<br />
}<br />
<br />
#clear {<br />
clear:both;<br />
}<br />
<br />
.shuttle_vec{<br />
width:362px;<br />
height:272px;<br />
}<br />
<br />
#shuttle_vec_cont{<br />
width:362px;<br />
height:390px;<br />
margin-left:300px;<br />
}<br />
<br />
</style></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:17:05Z<p>Sabrijamal: </p>
<hr />
<div>{{Team:Uppsala/repgenes-style}}<br />
{{Team:Uppsala/standard-style-no-frame}}<br />
<br />
<br />
<html><br />
<br />
<head><br />
<meta charset="utf-8" /> <br />
<title>Uppsala iGEM 2013</title> <br />
<link rel="shortcut icon" href="favicon.ico" type="image/x-icon"><br />
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<style type="text/css"><br />
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</head><br />
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<br />
<!--Main frame contains all the div elements--><br />
<br />
<div id= "main_frame"> <br />
<br />
<!--Contains our top header--><br />
<br />
<div id= "header"> <br />
<br />
<!-- Contains sub-logo picture --><br />
<div id="igem_logo"><a href="https://2013.igem.org/Team:Uppsala"><img class="igem_logo" src="https://static.igem.org/mediawiki/2013/e/e0/Uppsala2013_IGEMlogo.png"></a></div> <br />
<br />
<div id="igem"><a href="https://2013.igem.org/Main_Page"><img class="igem" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_Uppsala_IGEM_log_blue.png"></a></div><br />
<br />
<img class="slogan" src="https://static.igem.org/mediawiki/2013/a/ad/Uppsala2013_Header.png"> <br />
<br />
</div> <br />
<br />
<!-- Contains the "nav div" and the stripe background for the navigation menu --> <br />
<br />
<div id="navbar"> <br />
<br />
<!-- Contains the navigation menu with list elements, buged on local server, works on mediawiki!!--> <br />
<br />
<div id="nav"><br />
<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<div id="shuttle_vec_cont"> <img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img> <br />
<br />
<i> Above in the picture is a plate with our shuttle vector <a href="http://parts.igem.org/Part:BBa_K1033206">BBa_K1033206</a> containing the chromoprotein amilCP fused with the synthetic promoter CP29 from our promoter collection. The colonies are clearly blue efter 24 hours in E. coli, we have not yet been able to transform the construct to Lactobacillus. </i><br />
</div><br />
<br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
<br />
</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
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</html></div>Sabrijamalhttp://2013.igem.org/Template:Team:Uppsala/repgenes-styleTemplate:Team:Uppsala/repgenes-style2013-10-03T19:14:05Z<p>Sabrijamal: </p>
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</style></div>Sabrijamalhttp://2013.igem.org/Template:Team:Uppsala/repgenes-styleTemplate:Team:Uppsala/repgenes-style2013-10-03T19:13:41Z<p>Sabrijamal: </p>
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</style></div>Sabrijamalhttp://2013.igem.org/Template:Team:Uppsala/repgenes-styleTemplate:Team:Uppsala/repgenes-style2013-10-03T19:12:06Z<p>Sabrijamal: </p>
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<div><html><br />
<style type="text/css"><br />
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#main_frame {<br />
min-height: 1300px;<br />
width: 950px;<br />
margin: 20px auto;<br />
border: 1px solid ligth gray;<br />
border-radius: 15px;<br />
box-shadow: 0px 0px 30px rgba(110,110,110, .8); <br />
background: white;<br />
<br />
}<br />
<br />
.main-title {<br />
<br />
border:none;<br />
font-size:38px;<br />
color:#22409a;<br />
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}<br />
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<br />
border:none;<br />
font-size:38px;<br />
color:#22409a;<br />
float:left;<br />
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}<br />
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.second-row {<br />
border:none;<br />
font-size:38px;<br />
color:#22409a;<br />
position:relative;<br />
top:-20px;<br />
}<br />
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.no-border {<br />
border:none;<br />
}<br />
<br />
h1 {<br />
color:#0089d1;<br />
}<br />
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h3 {<br />
color:#0089d1;<br />
}<br />
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.symbol-pic {<br />
float:right;<br />
width:50px;<br />
height:50px;<br />
}<br />
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#clear {<br />
clear:both;<br />
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.shuttle_vec{<br />
width:362px;<br />
height:272px;<br />
margin-left:300px;<br />
}<br />
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#shuttle_vec_cont{<br />
width:362px;<br />
height:400px;<br />
border:solid 1px;<br />
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</style></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:11:07Z<p>Sabrijamal: </p>
<hr />
<div>{{Team:Uppsala/repgenes-style}}<br />
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<div id="igem_logo"><a href="https://2013.igem.org/Team:Uppsala"><img class="igem_logo" src="https://static.igem.org/mediawiki/2013/e/e0/Uppsala2013_IGEMlogo.png"></a></div> <br />
<br />
<div id="igem"><a href="https://2013.igem.org/Main_Page"><img class="igem" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_Uppsala_IGEM_log_blue.png"></a></div><br />
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<img class="slogan" src="https://static.igem.org/mediawiki/2013/a/ad/Uppsala2013_Header.png"> <br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<div id="shuttle_vec_cont"> <img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img> </div><br />
<br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
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</html></div>Sabrijamalhttp://2013.igem.org/Template:Team:Uppsala/repgenes-styleTemplate:Team:Uppsala/repgenes-style2013-10-03T19:09:42Z<p>Sabrijamal: </p>
<hr />
<div><html><br />
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</style></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:09:26Z<p>Sabrijamal: </p>
<hr />
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<div id="navbar"> <br />
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<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img><br />
<br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
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</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
<br />
</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:08:41Z<p>Sabrijamal: </p>
<hr />
<div>{{Team:Uppsala/repgenes-style}}<br />
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</div> <br />
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<!-- Contains the "nav div" and the stripe background for the navigation menu --> <br />
<br />
<div id="navbar"> <br />
<br />
<!-- Contains the navigation menu with list elements, buged on local server, works on mediawiki!!--> <br />
<br />
<div id="nav"><br />
<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
hej<br />
<img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img><br />
nej<br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
<br />
</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
<br />
</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:07:28Z<p>Sabrijamal: </p>
<hr />
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<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
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<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/1/18/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.png"> </img><br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
</div> <!-- main_ontent ends --> <br />
<br />
<div id="bottom-pic"><br />
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
</div><br />
<br />
<br />
<br />
</div> <!--Closing div element for main frame--><br />
<br />
<br />
<br />
</body><br />
<br />
</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:07:01Z<p>Sabrijamal: </p>
<hr />
<div>{{Team:Uppsala/repgenes-style}}<br />
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<title>Uppsala iGEM 2013</title> <br />
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<br />
<div id="navbar"> <br />
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<div id="nav"><br />
<div id="cow-box"><a href="https://2013.igem.org/Team:Uppsala"><img class="home-knapp" src="https://static.igem.org/mediawiki/2013/e/ef/Uppsala2013_Home.png"><a/></div><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
<br />
<br />
</ul></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li><br />
<br />
<li><a href="https://2013.igem.org/Team:Uppsala/team" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/a7/Uppsala2013_Team.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
<br />
</ul></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
</ul></li> <br />
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li><br />
</ul></li><br />
</ul><br />
</div> <br />
<br />
</div><br />
<br />
<div id="main_content"> <!-- Put content here --><br />
<br />
<h1 class="main-title-left">Reporter genes</h1><br />
<br />
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
<br />
<div id="clear"></div><br />
<br />
<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<img class="shuttle_vec" src="https://static.igem.org/mediawiki/2013/b/ba/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.JPG<br />
"> </img><br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
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</html></div>Sabrijamalhttp://2013.igem.org/Team:Uppsala/reporter-genesTeam:Uppsala/reporter-genes2013-10-03T19:06:45Z<p>Sabrijamal: </p>
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<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a><br />
<ul><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li><br />
</ul></li> <br />
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<li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a><br />
<ul><br />
<li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li><br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li> <br />
</ul></li><br />
</li> <br />
<li><a <br />
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">P-Coumaric acid</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li><br />
</ul></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li><br />
<li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li><br />
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<li><a href="https://2013.igem.org/Team:Uppsala/collaboration">Collaboration</a></li><br />
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<h1 class="main-title-left">Reporter genes</h1><br />
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<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/c/ca/Uppsala2013_Reporter_mini.png"><br />
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<p>One of the most fundamental parts required in a new chassi is a reporter gene. Unfortunately, some fluorescent proteins do not work well in Lactobacillus, possibly due to lower levels of internal pH in the cytosol.<br />
<br><br><br />
<p>One popular fluorescent protein that does work is mCherry. We have created a biobricked version of mCherry, <a href="http://parts.igem.org/Part:BBa_K1033250"> BBa_K1033250 </a> codon optimized for Lactobacillus reuteri, however it should work well in most species in the Lactobacillus genus. The gene has been modified in accordance with the Freiburgh fusion standard 25. Because of time constraints we have not yet had time to characterise it but it has been used in non-biobrick format by other researchers with positive results</p><br />
<br />
<h3>Prefix</h3><br />
<p>gaattccgcggccgcttctagatggccggc...</p><br />
<br />
<h3>Suffix</h3><br />
<p>... accggttaatactagtagcggccgctgcag</p><br />
<br />
<a id="b1"></a><br />
<h1>Chromoproteins in Lactobacillus </h1><br />
<p>There are many things we take for granted when working with bacteria such as E. coli. An initial problem when learning to work with Lactobacillus is to simply be able to do successful transformations of plasmids, something that is more difficult and cumbersome to do compared to E. coli. To see if you have been able to master a transformation with Lactobacillus some basic reporter genes that reflects visible light makes the validation process a lot easier.</p><br />
<br />
<img class="method-plasmid1" src="https://static.igem.org/mediawiki/2013/9/9b/Uppsala2013_reporter-plasmid.jpg"><br />
<br />
<p>Above is a picture of one of our chromo proteins and RBS fused together with two of our synthetic promoters, <a href="http://parts.igem.org/Part:BBa_K1033221"> CP11 </a> and <a href="http://parts.igem.org/Part:BBa_K1033221"> CP29 </a>. These promoters have been characterised and has been shown to work both in E. coli and Lactobacillus. We wanted to see if our chromoproteins could work as visual reporters. The construct has been shown to work fine with our shuttle vector in E. coli as can clearly be seen in the picture below with blue bacterial colonies. We have however not yet been able to transform it into Lactobacillus.</p><br />
<br />
<img class="shuttle_vec" src="ttp://2013.igem.org/wiki/images/b/ba/Uppsala2013_Shuttle_Vector_pSBLBC_cp29_amilCP1.JPG<br />
"> </img><br />
<br />
<h1> Biobricks </h1><br />
<br />
<a href="http://parts.igem.org/Part:BBa_K1033250">BBa_K1033250</a> - mCherry fusion protein codon optimised for Lactobacillus reuteri <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033280">BBa_K1033280</a> - aeBlue blue chromoprotein with strong promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033281">BBa_K1033281</a> - amilCP blue chromoprotein with medium strength promoter <br><br />
<a href="http://parts.igem.org/Part:BBa_K1033282">BBa_K1033282</a> - amilCP blue chromoprotein with strong promoter <br><br />
<br />
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<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png"><br />
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