Team:Toulouse/Project/binding

From 2014.igem.org

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<center><img style="width:700px; " src="http://2014.igem.org/wiki/images/e/e1/Bindingresume.png"></center>
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         <p class="texte"> In order to be highly efficient in the fight against the pythopathogen, our optimized bacterium has to be anchored to the fungus. Thus, we designed a chimeric protein (<a href="http://parts.igem.org/Part:BBa_K1364005"target="_blank">BBa_K1364005</a>) capable of building a <B>bridge between the bacterial peptidoglycan and the fungal chitin</B>, the main component of the pathogen’s cell wall. According to the work of the Imperial College 2010 iGEM team, we used the Cell Wall Binding (CWB) domain of the LytC protein (coding for a N-acetylmuramoyl-L-alanine amidase) to attach our chimeric protein to the <I>B. subtilis</I> cell wall. On the other side of our protein, we added the domain 4 of  GbpA from <I>Vibrio cholerae</I>, which is known to recognize chitin.
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         <p class="texte"> In order to be highly efficient in the fight against the pythopathogen, our optimized bacterium has to be anchored to the fungus. Thus, we designed a chimeric protein (<a href="http://parts.igem.org/Part:BBa_K1364005"target="_blank">BBa_K1364005</a>) capable of building a <B>bridge between the bacterial peptidoglycan and the fungal chitin</B>, the main component of the pathogen’s cell wall. According to the work of <a href="http://2010.igem.org/Team:Imperial_College_London"target="_blank">the Imperial College 2010</a> iGEM team, we used the Cell Wall Binding (CWB) domain of the LytC protein (coding for a N-acetylmuramoyl-L-alanine amidase) to attach our chimeric protein to the <I>B. subtilis</I> cell wall. On the other side of our protein, we added the domain 4 of  GbpA from <I>Vibrio cholerae</I>, which is known to recognize chitin.
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<br>The binding module has been placed under the control of Pveg (<a href="http://parts.igem.org/Part:BBa_K143012"target="_blank">BBa_K143012</a>), a strong constitutive promoter and we used a consensus RBS <a href="http://parts.igem.org/Part:BBa_K090505"target="_blank">BBa_K090505</a> and a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015"target="_blank">B0015</a>). The construct has been inserted in an integrative plasmid, pSBBS4S (<a href="http://parts.igem.org/Part:BBa_K823022"target="_blank">BBa_K823022</a>) which comes from the LMU-Munich 2012 iGEM team.</p>  
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<br>The binding module has been placed under the control of Pveg (<a href="http://parts.igem.org/Part:BBa_K143012"target="_blank">BBa_K143012</a>), a strong constitutive promoter and we used a consensus RBS (<a href="http://parts.igem.org/Part:BBa_K090505"target="_blank">BBa_K090505</a>) and a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015"target="_blank">B0015</a>). The construct has been inserted in an integrative plasmid, pSBBS4S (<a href="http://parts.igem.org/Part:BBa_K823022"target="_blank">BBa_K823022</a>) which comes from the LMU-Munich 2012 iGEM team.</p>  
<center style="margin:20px;"><img style="width:500px; " src="http://2014.igem.org/wiki/images/f/f5/BBa_K1364005.png"></center>
<center style="margin:20px;"><img style="width:500px; " src="http://2014.igem.org/wiki/images/f/f5/BBa_K1364005.png"></center>

Revision as of 11:51, 17 October 2014