Team:Marburg SYNMIKRO

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[[Image:Marburg_SYNMIKRO_team.png|Your team picture|right|frame]]
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== '''<html>&#160;</html>Recombinator:''' <br/> <html><sup>&#160;</sup></html>An intelligent Genetically Engineered Slot Machine (iGESM) ==
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<html>
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<div style="text-align:justify; padding-left:2px; padding-right:2px;">
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The vertebrate immune system produces billions of different antibodies. This diversity is generated by random VDJ-recombination of a limited number of antibody subfragments. This inspired us to construct an automatic recombination system in <i>E. coli</i> that generates large numbers of novel proteins by combinatorial fusion of functional domains. We used the site-specific DNA recombinase Gin of bacteriophage Mu that depends on the presence of a DNA enhancer element for efficient recombination. This allowed us to construct a system, called “The Recombinator”, which automatically shuts down after successful recombination. We visualized the randomizing function of our genetically engineered slot machine by combining fluorescent colors with cellular localization domains. By scaling up the number of recombination modules and functional domains our system will be able to generate a multitude of new proteins. We envision that “The Recombinator” will serve as a tool to create novel enzymatic activities for innovative drug design, environmental detoxification and metabolic engineering.
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<h3 style="z-index:1000;" >Team mixer</h3><a style="float:right; position: relative; top:-25px; z-index:1000;" href="https://2012.igem.org/Team:Marburg_SYNMIKRO/TeamProfile">[Team members]</a>
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== '''Overall project''' ==
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We aim to establish a system in Escherichia coli  that generates a large number of different proteins  by combinatorial fusion of few protein coding sub fragments.  
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    width: 163px;
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Our project was inspired by the VDJ-recombination of human immune system using a limited number of protein coding sequences to generate a huge diversity of antibodies.We designed an A and B module containing each three different protein coding sub fragments to be recombined. Recombination of one A fragment with one B fragment is achieved by the site-specific Gin recombinase of bacteriophage Mu. Specific Gix sites arranged as direct repeats are causing deletion within the modules and create combined fragments. Other Biobricks like a Gin enhancer and the suicide gene ccdb are located in between the modules and being deleted if an A and B fragment is fused. Those are necessary to ensure proper recombination. To visualize our results we decided to fuse fluorescent proteins of different color with intracellular localization domains. As proof of principle of our recombination machine, we expect E. colis to fluoresce in different colors at different cell domains.  
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(This might also give us the opportunity to quantify whether occurrence of curtain recombination products is related to the position of its sub fragments to the Gin enhancer)
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"<b>Daniel Kleinsorge</b><br><br>text",
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"<b>Oliver Schauer</b><br><br>text",
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"<b>Prof. Michael Bölker</b><br><br>text",
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"<b>Marian Vogt</b><br><br>text",
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"<b>Agnieszka Zarzycka</b><br><br>text",
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"<b>Oliver Leicht</b><br><br>text",
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"<b>Margarita Derbenev</b><br><br>text",
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"<b>Elisabeth Ignatz</b><br><br>text",
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"<b>Julian Pechstein</b><br><br>text",
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"<b>Johannes Schuh</b><br><br>text",
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"<b>Max Diesner</b><br><br>text",
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"<b>Svetlana Uscherenko</b><br><br>text"
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);
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<div>
<br>
<br>
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<a onclick="random(); " class="btn"><span class="blink">Recombine!</span></a>
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</center>
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</div>       
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</div>
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</div>
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</div>
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<a href="https://twitter.com/iGESM_Marburg" class="twitter-follow-button" data-show-count="false" data-lang="de">@iGESM_Marburg folgen</a>
 
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<html>
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<div  style="top:345px; position: absolute; width:61.5%; height:299px;" class="conbox60">
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</html>
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== Bacterial combinatorics ==
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As proof-of-principle of our recombinator system we created combinatorial fusions of bacterial intracellular localization domains with different fluorescent proteins. We expected to generate a mixture of bacteria that glow in different colors and at different positions in all possible combinations.
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<br/>
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<br/>
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[[File:Syn_MR_bunte_Maus.png|center|420px]]
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Here, we have dealt only with few modules, but with additional recombination modules, the number of possible combinations raises exponentially.
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<html>
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</div>
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<div class="conboxb">
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</html>
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[[File:Syn_MR_Logo.png|link=http://www.uni-marburg.de|left|400px]][[File:Syn_MR_synmikro_farbe.gif|link=http://www.synmikro.com/en/|right|400px]]
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<html>
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</div>
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Latest revision as of 13:25, 23 September 2013

Main Page

 Recombinator:
 An intelligent Genetically Engineered Slot Machine (iGESM)

The vertebrate immune system produces billions of different antibodies. This diversity is generated by random VDJ-recombination of a limited number of antibody subfragments. This inspired us to construct an automatic recombination system in E. coli that generates large numbers of novel proteins by combinatorial fusion of functional domains. We used the site-specific DNA recombinase Gin of bacteriophage Mu that depends on the presence of a DNA enhancer element for efficient recombination. This allowed us to construct a system, called “The Recombinator”, which automatically shuts down after successful recombination. We visualized the randomizing function of our genetically engineered slot machine by combining fluorescent colors with cellular localization domains. By scaling up the number of recombination modules and functional domains our system will be able to generate a multitude of new proteins. We envision that “The Recombinator” will serve as a tool to create novel enzymatic activities for innovative drug design, environmental detoxification and metabolic engineering.

Team mixer

[Team members]

Bacterial combinatorics

As proof-of-principle of our recombinator system we created combinatorial fusions of bacterial intracellular localization domains with different fluorescent proteins. We expected to generate a mixture of bacteria that glow in different colors and at different positions in all possible combinations.

Syn MR bunte Maus.png

Here, we have dealt only with few modules, but with additional recombination modules, the number of possible combinations raises exponentially.

Syn MR Logo.png
400px

Retrieved from "http://2012.igem.org/Team:Marburg_SYNMIKRO"