Team:Toulouse/Project/Chemotaxis

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<center><img style="width:420px; " src="https://static.igem.org/mediawiki/parts/e/e9/Recap_chemotax.jpg"></center>
<center><img style="width:420px; " src="https://static.igem.org/mediawiki/parts/e/e9/Recap_chemotax.jpg"></center>
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<p class="legend">Figure 1: Schema of the chemotaxis module</p>
<p class="title1">What is chemotaxis?</p>
<p class="title1">What is chemotaxis?</p>
<p class="texte">
<p class="texte">
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Chemotaxis is a bacterial function which allows to move according to a concentration gradient. With this system bacteria can find  a better place to grow by swimming toward higher concentrations of molecules, such as nutritional molecules like sugar, amino acid, vitamins...Chemotactic-signal transducers respond to changes in the concentration of attractants and repellents in the environment, transduce a signal from the outside to the inside of the cell, and facilitate sensory adaptation through the variation of the level of methylation.  
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Chemotaxis is a bacterial function which induces a movement toward a gradient of concentration of a molecule of interest. With this system the bacteria are able to swim to a location containing higher concentrations of molecules such as sugar, amino acid, vitamins...  
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Chemotactic-signal transducers respond to changes in the concentration of attractants and repellents in the environment, transduce the signal from the outside to the inside of the cell, and facilitate sensory adaptation through the variation of the level of methylation.  
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<p class="title1">More information on this module</p>
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In our case, chemotaxis is used as a way to detect and approach fungi. Indeed during its growth, fungi release N-acetylglucosamine (NAG), the basic unit of chitin which composed its cell wall. Thus, the concentration of N-acetylglucosamine is getting more important around fungi.</p>
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<p class="texte">
<p class="texte">
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It is well known that <i>Bacillus subtilis</i> is able to detect and to shift towards glucose thanks to the Methyl-accepting chemotaxis protein, called <b> McpA </b>. <br>
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Chemotaxis is used as a way to detect and come close to the location of fungi infection. During its growth, fungi release N-acetylglucosamine (NAG), the basic unit of chitin which composed its cell wall. Thus, there should exist a gradient of the concentration of NAG around the fungi.</p>
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Some bacteria are attracted by NAG, like <i>Vibrio cholerae</i> which has a N-acetylglucosamine regulated methyl-accepting chemotaxis protein, henceforth called with the simple name of<b> VCD</b>.</p>
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<p class="texte">
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It is known that <i>B. subtilis</i> is able to detect and to swim towards glucose using the Methyl-accepting chemotaxis protein, henceforth called <b>McpA</b> (<a href="http://www.uniprot.org/uniprot/P39214"_blanck">MCPA_BACSU</a>).<br>
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Some bacteria are attracted by NAG, like <i>Vibrio cholerae</i> which has a NAG regulated methyl-accepting chemotaxis protein: <b>VCD</b> (<a href="http://www.uniprot.org/uniprot/C3NYT2"_blank">VCD_000306</a>).</p>
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<center><img SRC="https://static.igem.org/mediawiki/2014/c/cf/Schema_chemotaxis.png" alt="schema"></center>
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<center><img width="500px" SRC="https://static.igem.org/mediawiki/2014/4/47/Chimio1.png" alt="schema" style="margin-bottom:60px;"></center>
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<p class="legend">Figure 2: Chimeric protein of chemotaxis</p>
<p class="texte">
<p class="texte">
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Therefore, our idea is to switch the natural glucose specificity of <i>B. subtilis</i> by a NAG specificity. To achieve this, we need to change the extracellular domain of McpA, the domain responsible for the specificity, by the extracellular domain of VCD.
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Therefore, our idea is to switch the natural glucose specificity of <i>B. subtilis'</i>, mediated by McpA, to a NAG specificity. To achieve this, we need to change the extracellular domain of McpA, responsible for the specificity, by the extracellular domain of VCD.
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The whole sequence has been designed <i>in silico</i> and codon optimized for the transcription in <i>Bacillus subtilis</i> before its synthesis.</p>
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The whole sequence has been designed <i>in silico</i> and codon optimized for the transcription in <i>B. subtilis</i> before its synthesis.</p>
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<center><img SRC="https://static.igem.org/mediawiki/2014/c/cf/Gene_chemotaxis.png" alt="gene construct"></center>
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<center><img width="600px" SRC="https://static.igem.org/mediawiki/2014/e/e4/Chimio2.png" alt="gene construct" style="margin-bottom:40px;"></center>
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<p class="legend">Figure 3: Construction of the chemotaxis gene</p>
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<p class="title1">References</p>
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<center><a href="https://2014.igem.org/Team:Toulouse/Result/experimental-results"> <img src="https://static.igem.org/mediawiki/parts/f/fe/Jump.jpg"> </a></center>
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<p class="texte">
 
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- K. Meibom,L. Xibing, A. Nielsen, CY. Wu, S. Roseman and G. Schoolnik.<b> The Vibrio cholerae chitin utilization program </b>. The National Academy of Sciences of the USA (2004). <br>
 
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-C. Kristich and GW. Ordal.<b>Bacillus subtilis CheD is a chemoreceptor modification enzyme required for chemotaxis</b>. J Biol Chem. 2002 Jul 12;277(28):25356-62. Epub 2002 May 13.<br></p>
 
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<p class="title1">References</p>
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<ul>
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<li class="tree"><p class="texte">K. Meibom,L. Xibing, A. Nielsen, CY. Wu, S. Roseman, and G. Schoolnik.<b> The <i>Vibrio cholerae</i> chitin utilization program </b>. The National Academy of Sciences of the USA (2004).</p></li>
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<li class="tree"><p class="texte">C. Kristich, and GW. Ordal. <b><i>Bacillus subtilis</i> CheD is a chemoreceptor modification enzyme required for chemotaxis</b>. J Biol Chem. 2002 Jul 12;277(28):25356-62. Epub 2002 May 13.<br></p></li>
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</ul>
      
      
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Latest revision as of 02:59, 18 October 2014