Team:iHKU/design

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          <td width="50%"><span class="style3"><a href="http://www.hku.hk/">The University of Hong Kong</a> | <a href="http://www.hku.hk/facmed/">The Faculty of Medicine</a></span></td>
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                        <td width="80%" align="left"><h1 class="style7">Design</h1>
                          <h3 class="style7">1. 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>
                          <h3><strong>2. 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>(Diagrams &amp; picture to be added)</p>
                          <h3><strong>3. 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 &amp;  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>
                          <p align="center"><img src="design_pic1.png" width="525" height="352" /></p>
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                          <p>Predicted Pattern:</p>
                          <h1 align="center"><img src="image/design_predicted.png" width="465" height="239" /></h1>
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                          <h1>Plasmids and strains</h1>
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