Team:iHKU/design

From 2008.igem.org

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                       <th width="10%">&nbsp;</th>
                       <th width="10%">&nbsp;</th>
                         <td width="80%" align="left"><h1 class="style7">Design</h1>
                         <td width="80%" align="left"><h1 class="style7">Design</h1>
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                           <h3 class="style7">Our Aim</h3>
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                          <ul>
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                            <li class="style18"><strong><a href="#1"><span class="style7">Our Aim</span></a></strong></li>
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                            <li class="style18"><strong><a href="#2">Chassis  selection</a></strong></li>
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                            <li class="style18"><strong><a href="#3">Genetic  Circuit Design</a></strong></li>
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                            <li class="style18"><strong><a href="#4">Plasmids and strains</a></strong></li>
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                          </ul>
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                           <h3 class="style7">&nbsp;</h3>
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                          <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>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>
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                           <h3><strong>Chassis  selection</strong></h3>
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                           <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>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></p>
                           <p></p>
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                           <h3><strong>Genetic  Circuit Design</strong></h3>
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                           <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 &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 />
                           <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>
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>
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                           <h1 align="center"><img src="/wiki/images/thumb/1/1e/Design_pic3.png/800px-Design_pic3.png" width="465" height="230" /></h1>
                           <p>&nbsp;</p>
                           <p>&nbsp;</p>
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                           <h3><strong>Plasmids and strains</strong></h3>
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                           <h3><strong><a name="4" id="4"></a>Plasmids and strains</strong></h3>
                         </td>
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Revision as of 16:26, 29 October 2008

 

Design

 

Our Aim

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.

Chassis selection

Past chemotaxis studies have provided the molecular basis of cellular motility regulation, Escherichia coli and Bacillus subtilis are notably the well-understood strains which are ideal to be the chassis of our designed genetic circuit. We chose E.coli as our chassis for the project reasoning that cell-cell communications will require the use of a signaling molecule, which are often density related. E.coli is known to be less motile than Bacillus in terms of speed, thus would ease the accumulation of the signaling molecule. We hope the subsequent pattern generated by using E.coli as chassis would be finer and more interesting.

Genetic Circuit Design

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 cheZgene, and can be restored by subsequent re-introduction of the cheZ gene under a controllable promoter back into the host.
We designed two DNA constructs whose cheZ expression level would be sensitive to the concentration of AHL (Acetyl homoserine lactone). One would become motile in the presence of AHL, while the other one be become immotile under the same condition. Since concentration of AHL is proportional to cell density, the motility of our strains would be dependent on local cell density.

 

Predicted Pattern:

 

Plasmids and strains

  
 

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