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Team:iHKU/design
From 2008.igem.org
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| - | <td width=" | + | <td width="39%"><span class="headline"><a href="http://www.hku.hk">The University of Hong Kong</a> | <a href="http://www.hku.hk/facmed/">Li Ka Shing Faculty of Medicine</a></span></td> |
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<td width="80%" align="left"><h1 class="style7">Design</h1> | <td width="80%" align="left"><h1 class="style7">Design</h1> | ||
<ul> | <ul> | ||
| - | <li class="style18" | + | <li class="style18"><a href="#1">Our Aim</a></u></li> |
| - | <li class="style18" | + | <li class="style18"><a href="#2">Chassis selection</a></li> |
| - | <li class="style18" | + | <li class="style18"><a href="#3">Genetic Circuit Design</a></li> |
| - | <li class="style18" | + | <li class="style18"><a href="#4">Plasmids and strains</a></li> |
</ul> | </ul> | ||
<h3 class="style7"> </h3> | <h3 class="style7"> </h3> | ||
<h3 class="style7"><strong><a name="1" id="1"></a></strong>Our Aim</h3> | <h3 class="style7"><strong><a name="1" id="1"></a></strong>Our Aim</h3> | ||
| - | <p>We endeavor our strains to grow into patterns by arranging themselves in a synchronous, self-organised manner, “just as in organisms in nature which all are able to develop shapes and patterns.” Implementing such idea on bacteria sheds light to a mechanism involving cell-cell communication that would produce a key response, predominately a respond affecting cell motility. The characteristics of the response logically should be critical towards the formation overall pattern. </p> | + | <p class="special">We endeavor our strains to grow into patterns by arranging themselves in a synchronous, self-organised manner, “just as in organisms in nature which all are able to develop shapes and patterns.” Implementing such idea on bacteria sheds light to a mechanism involving cell-cell communication that would produce a key response, predominately a respond affecting cell motility. The characteristics of the response logically should be critical towards the formation overall pattern. </p> |
<p align="right"><a href="#top">[Back to Top]</a></p> | <p align="right"><a href="#top">[Back to Top]</a></p> | ||
<h3><strong><a name="2" id="2"></a>Chassis selection</strong></h3> | <h3><strong><a name="2" id="2"></a>Chassis selection</strong></h3> | ||
| - | <p>Past chemotaxis studies have provided the molecular basis of cellular motility regulation, <em>Escherichia coli</em> and Bacillus subtilis are notably the well-understood strains which are ideal to be the chassis of our designed genetic circuit. We chose <em>E.coli</em> as our chassis for the project reasoning that cell-cell communications will require the use of a signaling molecule, which are often density related. <em>E.coli</em> is known to be less motile than <em>Bacillus</em> in terms of speed, thus would ease the accumulation of the signaling molecule. We hope the subsequent pattern generated by using <em>E.coli</em> as chassis would be finer and more interesting.</p> | + | <p class="special">Past chemotaxis studies have provided the molecular basis of cellular motility regulation, <em>Escherichia coli</em> and Bacillus subtilis are notably the well-understood strains which are ideal to be the chassis of our designed genetic circuit. We chose <em>E.coli</em> as our chassis for the project reasoning that cell-cell communications will require the use of a signaling molecule, which are often density related. <em>E.coli</em> is known to be less motile than <em>Bacillus</em> in terms of speed, thus would ease the accumulation of the signaling molecule. We hope the subsequent pattern generated by using <em>E.coli</em> as chassis would be finer and more interesting.</p> |
<p align="right"><a href="#top">[Back to Top]</a></p> | <p align="right"><a href="#top">[Back to Top]</a></p> | ||
<p></p> | <p></p> | ||
<h3><strong><a name="3" id="3"></a>Genetic Circuit Design</strong></h3> | <h3><strong><a name="3" id="3"></a>Genetic Circuit Design</strong></h3> | ||
| - | <p>An On-Off motility design is desired as the response to cell-cell communication. Based on the pioneer work (Topp & Gallivan 2006) shows the motility can be abolished by knocking out the <strong><em>cheZ</em></strong>gene, and can be restored by subsequent re-introduction of the <em>cheZ</em> gene under a controllable promoter back into the host.<br /> | + | <p class="special">An On-Off motility design is desired as the response to cell-cell communication. Based on the pioneer work (Topp & Gallivan 2006) shows the motility can be abolished by knocking out the <strong><em>cheZ</em></strong>gene, and can be restored by subsequent re-introduction of the <em>cheZ</em> gene under a controllable promoter back into the host.<br /> |
We designed two DNA constructs whose cheZ expression level would be sensitive to the concentration of AHL (Acetyl homoserine lactone). One would become <strong>motile</strong> in the presence of AHL, while the other one be become <strong>immotile </strong>under the same condition. Since concentration of AHL is proportional to <strong>cell density</strong>, the motility of our strains would be dependent on local cell density.</p> | We designed two DNA constructs whose cheZ expression level would be sensitive to the concentration of AHL (Acetyl homoserine lactone). One would become <strong>motile</strong> in the presence of AHL, while the other one be become <strong>immotile </strong>under the same condition. Since concentration of AHL is proportional to <strong>cell density</strong>, the motility of our strains would be dependent on local cell density.</p> | ||
<p align="center"><img src="/wiki/images/4/4f/Lux.gif" width="565" height="175" /></p> | <p align="center"><img src="/wiki/images/4/4f/Lux.gif" width="565" height="175" /></p> | ||
Revision as of 17:23, 29 October 2008
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