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Team:Toulouse/Project/Chemotaxis
From 2014.igem.org
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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> | ||
| + | <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"> | ||
| - | Chemotaxis is a bacterial function which | + | 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... |
| + | 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. | ||
<p class="title1">More information on this module</p> | <p class="title1">More information on this module</p> | ||
<p class="texte"> | <p class="texte"> | ||
| - | Chemotaxis is used as a way to detect and | + | 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> |
<p class="texte"> | <p class="texte"> | ||
| - | It is | + | 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> |
| - | + | 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> | |
<center><img width="500px" SRC="https://static.igem.org/mediawiki/2014/4/47/Chimio1.png" alt="schema" style="margin-bottom:60px;"></center> | <center><img width="500px" SRC="https://static.igem.org/mediawiki/2014/4/47/Chimio1.png" alt="schema" style="margin-bottom:60px;"></center> | ||
| + | <p class="legend">Figure 2: Chimeric protein of chemotaxis</p> | ||
| + | |||
<p class="texte"> | <p class="texte"> | ||
| - | 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, | + | 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. |
| - | The whole sequence has been designed <i>in silico</i> and codon optimized for the transcription in <i> | + | 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> |
<center><img width="600px" SRC="https://static.igem.org/mediawiki/2014/e/e4/Chimio2.png" alt="gene construct" style="margin-bottom:40px;"></center> | <center><img width="600px" SRC="https://static.igem.org/mediawiki/2014/e/e4/Chimio2.png" alt="gene construct" style="margin-bottom:40px;"></center> | ||
| + | <p class="legend">Figure 3: Construction of the chemotaxis gene</p> | ||
| + | |||
| + | <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> | ||
| + | |||
<p class="title1">References</p> | <p class="title1">References</p> | ||
<ul> | <ul> | ||
| - | <li class="tree"><p class="texte">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).</p></li> | + | <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> |
| - | <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> | + | <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> |
</ul> | </ul> | ||
Latest revision as of 02:59, 18 October 2014
Chemotaxis
To target the pathogenic fungus
Project > Chemotaxis
Figure 1: Schema of the chemotaxis module
What is chemotaxis?
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... 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.
More information on this module
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.
It is known that B. subtilis is able to detect and to swim towards glucose using the Methyl-accepting chemotaxis protein, henceforth called McpA (MCPA_BACSU).
Some bacteria are attracted by NAG, like Vibrio cholerae which has a NAG regulated methyl-accepting chemotaxis protein: VCD (VCD_000306).
Figure 2: Chimeric protein of chemotaxis
Therefore, our idea is to switch the natural glucose specificity of B. subtilis', 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. The whole sequence has been designed in silico and codon optimized for the transcription in B. subtilis before its synthesis.
Figure 3: Construction of the chemotaxis gene
References
K. Meibom,L. Xibing, A. Nielsen, CY. Wu, S. Roseman, and G. Schoolnik. The Vibrio cholerae chitin utilization program . The National Academy of Sciences of the USA (2004).
C. Kristich, and GW. Ordal. Bacillus subtilis CheD is a chemoreceptor modification enzyme required for chemotaxis. J Biol Chem. 2002 Jul 12;277(28):25356-62. Epub 2002 May 13.
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