Team:Toulouse/Project/binding

From 2014.igem.org

(Difference between revisions)
Line 85: Line 85:
<center><img style="width:700px; " src="http://2014.igem.org/wiki/images/e/e1/Bindingresume.png"></center>
<center><img style="width:700px; " src="http://2014.igem.org/wiki/images/e/e1/Bindingresume.png"></center>
-
         <p class="texte"> In order to be more effective in the fight against the pythopathogen, our optimized bacterium has to be anchored to the fungus. This module matches with the <B> binding ability </B> of SubtiTree. Thus, we designed a chimeric protein (<a href="http://parts.igem.org/Part:BBa_K1364005"target="_blank">BBa_K1364005</a>) to make <B> a bridge between the bacterial peptidoglycan and the fungal chitin </B>, which is the main component of the pathogen’s cell wall. According to the work of the 2010 Imperial College iGEM team, we used the CWB domain of the LytC protein (coding for a N-acetylmuramoyl-Lalanine amidase) to bind our chimeric protein to <I> Bacillus subtilis' </I> cell wall. On the other side of our protein, we added the 4th domain of  GbpA from <I> Vibrio cholerae </I>, known to recognize N-AcetylGlucosamine oligosaccharides called chitin.
+
         <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 <B>a 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 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>
Line 94: Line 94:
<ul>
<ul>
-
<li class="tree"><p class="texte"><B>Anchor section </B>: the CWB (Cell Wall Binding) domain is extracted from LytC gene and composed the 5' side of our binding module. As previously used by the 2010 Imperial College of London iGEM team, we kept the first 318 bp. We can note the presence of the signal peptide at the beginning of the sequence, from 1 to 24 bp.</p></li>
+
<li class="tree"><p class="texte"><B>Anchor section</B>: the CWB (Cell Wall Binding) domain is extracted from LytC gene and composes the 5' side of our binding module. As previously described by the Imperial College of London 2010  iGEM team, we kept the first 318 bp. We can note the presence of the signal peptide at the beginning of the sequence, from 1 to 24 bp.</p></li>
-
<li class="tree"><p class="texte"><B> Chitin Binding Domain (CBD) section </b>:  the 4th domain of GbpA from <I>Vibrio cholerae </I> is able to bind to N-Acetyl Glucosamine oligosacchararides. Also,  the 3' side of our gene is composed by a part of the GbpA sequence (from 423 to 484 bp).</p></li>
+
<li class="tree"><p class="texte"><B>Chitin Binding Domain (CBD) section</b>:  the 4th domain of GbpA from <I>Vibrio cholerae </I> is able to bind to N-Acetyl Glucosamine oligosacchararides. Also,  the 3' side of our gene is composed by a part of the GbpA sequence (from 423 to 484 bp).</p></li>
-
<li class="tree"><p class="texte"><B>Helical Linker </B>: According to the work of the 2010 Imperial College of London iGEM team, we used the same six amino acids sequence (SRGSRA) to make a bridge between the Anchor section and the Chitin Binding section.
+
<li class="tree"><p class="texte"><B>Helical Linker</B>: According to the work of the 2010 Imperial College of London iGEM team, we used the same six amino acids sequence (SRGSRA) to make a bridge between the Anchor section and the Chitin Binding section.
</p></li></ul>
</p></li></ul>
<center style="margin:-30px;"><img style="width:500px; " src="http://2014.igem.org/wiki/images/a/a0/Binding.png"></center>
<center style="margin:-30px;"><img style="width:500px; " src="http://2014.igem.org/wiki/images/a/a0/Binding.png"></center>
Line 109: Line 109:
<p class="texte">
<p class="texte">
-
<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>  
+
<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 15:29, 16 October 2014