Team:Toulouse/Project/binding

From 2014.igem.org

(Difference between revisions)
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     <div class="banniere-content">
     <div class="banniere-content">
       <h2>Binding</h2>
       <h2>Binding</h2>
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       <p>To be attached to the fungal wall</p>
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       <p>To be attached to the fungal cell wall</p>
     </div>
     </div>
   </div>
   </div>
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<p class="legend">Figure 1: Schema of the binding module</p>
<p class="legend">Figure 1: Schema of the binding module</p>
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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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         <p class="texte"> In order to be highly efficient in the fight against the pythopathogen <i>Ceratocystis platani</i>, our optimized bacterium has to be anchored to the fungus.  
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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.
</p>
</p>
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<p class="title1"> More information about this module </p>
<p class="title1"> More information about this module </p>
<p class="texte">  
<p class="texte">  
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The Binding Module ORF is composed of 3 sections:</p>
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The open reading frame of the Binding Module is composed of 3 sections:</p>
<ul>
<ul>
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<p class="texte">
<p class="texte">
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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>),
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a strong constitutive promoter and we used a consensus RBS (<a href="http://parts.igem.org/Part:BBa_K090505"target="_blank">BBa_K090505</a>) as well as a
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double terminator (<a href="http://parts.igem.org/Part:BBa_B0015"target="_blank">B0015</a>).  
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</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 20:27, 17 October 2014