Team:Uppsala/affinity-tags

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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/2013/9/9a/Home.png"><a/></div>
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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>
<ul>
<ul>
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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/2013/9/92/Project.png"></a>
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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>
<ul>
<ul>
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<li><a href="https://2013.igem.org/Team:Uppsala/overview">Overwiev</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/overview">Overview</a></li>
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<li><a href="#">Chassi</a>
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a>
<ul>
<ul>
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                                                                <li><a href="https://2013.igem.org/Team:Uppsala/probiotics">Probiotics</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/affinity-tags">Affinity tags</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/affinity-tags">Toxin-antitoxin system</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li>
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                                                                <li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li>
</ul></li>
</ul></li>
                                                  
                                                  
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                                                 <li><a href="#">Metabolic engineering</a>
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                                                 <li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a>
                                                     <ul>
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                                                                 <li><a href="#">Resveratrol</a></li>
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                                                                 <li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li>
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                                                                <li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/beta-carotene">β-carotene</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/betacarotene">β-carotene</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/astraxantin">Astraxantin</a></li>
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                                                                <li><a
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<li><a href="https://2013.igem.org/Team:Uppsala/zeaxantin">Zeaxantin</a></li>
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href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/astraxantin">Astaxanthin</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/zeaxantin">Zeaxanthin</a></li>
</ul></li>
</ul></li>
</li>  
</li>  
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<li><a href="#">Chromoprotein</a></li>
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                                                <li><a
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<li><a href="#">Safety experiments</a></li>
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href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li>
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<li><a href="#">Result</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/safety">Safety experiment</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/result">Result</a></li>
</ul></li>
</ul></li>
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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/2013/3/3c/Modeling.png"></a>
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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>
<ul>
<ul>
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<li><a href="#">Resveratrol pathway </a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-pathway">Resveratrol pathway </a></li>
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<li><a href="#">Carotenoids pathway </a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li>
</ul></li>
</ul></li>
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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/2013/7/79/Parts.png"></a></li>
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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>
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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/2013/3/32/Team.png"></a>
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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>
<ul>
<ul>
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<li><a href="#">Resveratrol group</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-group">Resveratrol group</a></li>
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<li><a href="#">Carotenoid group</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li>
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<li><a href="#">Chassi group</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li>
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<li><a href="#">Advisors</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/advisors">Advisors</a></li>
</ul></li>
</ul></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/hp" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/f/f5/HP.png"></a>
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<li><a href="https://2013.igem.org/Team:Uppsala/hp" id="list_type3"><img class="nav-text"       src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a>
<ul>
<ul>
<li><a href="https://2013.igem.org/Team:Uppsala/Yoghurtproducts">Yoghurt products</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/Yoghurtproducts">Yoghurt products</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">Synbioday</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBio day</a></li>
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<li><a href="#">Bioethics</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/safety">Bioethics and safety</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/safety">Society & Safety</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/society">Society and outreach</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/society">Society</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/highschool-outreach">High school outreach</a></li>
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<li><a href="https://2013.igem.org/Team:Uppsala/hsoutreach">High School Outreach</a></li>
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                                                <li><a href="https://2013.igem.org/Team:Uppsala/outreach">Media outreach</a></li>
</ul></li>
</ul></li>
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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/2013/8/8d/Attributions.png"></a></li>  
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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>  
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<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/9/95/Notebook.png"></a></li>
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<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>
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                                    <ul>
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                                        <li><a href="https://2013.igem.org/Team:Uppsala/protocol">Protocol</a></li>
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                                        <li><a href="https://2013.igem.org/Team:Uppsala/notebook">Lab journal</a></li>
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                                    </ul></li>
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<h1 class="main-title-left"> Toxin-antitoxin system </h1>
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<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/1/12/Uppsala2013_toxin-antitoxin_symbol_mini.png">
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<h1> Keep your plasmids without antibiotic resistance </h1>
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One of the challenges when creating synthetic systems in bacteria that serve a purpose besides increasing the fitness of the organism is that there is a negative selective pressure against keeping the system. Toxin-antitoxin systems can be used to make plasmids far more stabile without having to use antibiotics and antibiotic resistance.  If a clone were to lose the plasmid, the toxin which usually has a longer half life than the antitoxin will kill the bacteria.
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<img class="method-plasmid" src="https://static.igem.org/mediawiki/2013/d/dc/Uppsala2013_anti-toxin-toxin-system.jpg">
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<p>Above is an example of how a toxin-antitoxin system could be applied together with a gene X. If the gene is toxic or expressed strongly enough there will be a substantial evolutionary pressure to lose the plasmid during cell division. However if the gene is present on a plasmid with a toxin-antitoxin system would be lethal due to the loss of the antitoxin gene.</p>
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<h1> A natural toxin-antitoxin from lactobacillus plantarum </h1>
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We have taken the Pem toxin-antitoxin system from plasmid p256 that was originally isolated from lactobacillus plantarum NC7. The system consists of a single operon and consists of two ORFs, one for the toxin and antitoxin respectively. Pem on p256 has been shown to increase segregational stability under non-selective pressure. The system has experimentally been shown to allow 88-100% retention of a plasmid after 80 generations(1). We have provided the toxin-antitoxin system both with and without a natural putative promoter.
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<br>
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<h1> References: </h1>
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http://mic.sgmjournals.org/content/151/2/421.long (1)

Latest revision as of 05:42, 29 September 2013

Uppsala iGEM 2013

Toxin-antitoxin system

Keep your plasmids without antibiotic resistance

One of the challenges when creating synthetic systems in bacteria that serve a purpose besides increasing the fitness of the organism is that there is a negative selective pressure against keeping the system. Toxin-antitoxin systems can be used to make plasmids far more stabile without having to use antibiotics and antibiotic resistance. If a clone were to lose the plasmid, the toxin which usually has a longer half life than the antitoxin will kill the bacteria.

Above is an example of how a toxin-antitoxin system could be applied together with a gene X. If the gene is toxic or expressed strongly enough there will be a substantial evolutionary pressure to lose the plasmid during cell division. However if the gene is present on a plasmid with a toxin-antitoxin system would be lethal due to the loss of the antitoxin gene.

A natural toxin-antitoxin from lactobacillus plantarum

We have taken the Pem toxin-antitoxin system from plasmid p256 that was originally isolated from lactobacillus plantarum NC7. The system consists of a single operon and consists of two ORFs, one for the toxin and antitoxin respectively. Pem on p256 has been shown to increase segregational stability under non-selective pressure. The system has experimentally been shown to allow 88-100% retention of a plasmid after 80 generations(1). We have provided the toxin-antitoxin system both with and without a natural putative promoter.

References:

http://mic.sgmjournals.org/content/151/2/421.long (1)

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