http://2012.igem.org/wiki/index.php?title=Team:XMU-China/timedelay&feed=atom&action=historyTeam:XMU-China/timedelay - Revision history2024-03-28T16:45:17ZRevision history for this page on the wikiMediaWiki 1.16.0http://2012.igem.org/wiki/index.php?title=Team:XMU-China/timedelay&diff=296679&oldid=prevIcedcat at 02:57, 27 October 20122012-10-27T02:57:41Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>Figure 4:</b> The optimization of the arabinose concentration added into the TD1.0 including<br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>Figure 4:</b> The optimization of the arabinose concentration added into the TD1.0 including<br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>0, 0.1, 1.0, 10 mM arabinose.<a target="_blank" href=" https://static.igem.org/mediawiki/2012/e/e4/-Fig_4-Optimization_of_arabinose_induced_concentration_%28TD1.0%29.jpg">[click to data]</a></p><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>0, 0.1, 1.0, 10 mM arabinose.<a target="_blank" href=" https://static.igem.org/mediawiki/2012/e/e4/-Fig_4-Optimization_of_arabinose_induced_concentration_%28TD1.0%29.jpg">[click to data]</a></p><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p align="left">We can find that the curve with 0.<del class="diffchange diffchange-inline">1mM </del>arabinose has a higher fluorescence. So we chose the </a>0.1 mM arabinose for the following fluorescence tests.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p align="left">We can find that the curve with 0.<ins class="diffchange diffchange-inline">1 mM </ins>arabinose has a higher fluorescence. So we chose the </a>0.1 mM arabinose for the following fluorescence tests.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p class="subsubtitle">4.2 Fluorescent test of the time delay devices</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p class="subsubtitle">4.2 Fluorescent test of the time delay devices</p></div></td></tr>
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</table>Icedcathttp://2012.igem.org/wiki/index.php?title=Team:XMU-China/timedelay&diff=287516&oldid=prevYoubin at 18:13, 26 October 20122012-10-26T18:13:54Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 1:</b> The mechanism graph of iccdB last year<sup><a href="#_ENREF_3" title="Ron Weiss, 2003 #2">[3]</a></sup></p><hr><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 1:</b> The mechanism graph of iccdB last year<sup><a href="#_ENREF_3" title="Ron Weiss, 2003 #2">[3]</a></sup></p><hr><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="subtitle"><a name="_Toc03" id="Toc03"></a>3. Constructions this year</span></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="subtitle"><a name="_Toc03" id="Toc03"></a>3. Constructions this year</span></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/<del class="diffchange diffchange-inline">5</del>/<del class="diffchange diffchange-inline">57</del>/<del class="diffchange diffchange-inline">Timedelay_f2reversed</del>.<del class="diffchange diffchange-inline">png</del>" alt="" width="420" /><br /></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/<ins class="diffchange diffchange-inline">e</ins>/<ins class="diffchange diffchange-inline">e6</ins>/<ins class="diffchange diffchange-inline">Delay_01</ins>.<ins class="diffchange diffchange-inline">jpg</ins>" alt="" width="420" /><br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 2:</b> The construction devices of time delay system<br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 2:</b> The construction devices of time delay system<br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p align="left">Based on our last year&rsquo;s project, for convenience of testing the results of time delay device, we chose GFP instead of CcdB as our reporter. On the other hand, because of the basal expression of <em>P<sub>lac</sub></em> is very strong, we decided to use arabinose to induce <em>P<sub>BAD</sub></em> promoter to activate our device.<sup><a href="#_ENREF_4" title="Ron Weiss, 2003 #2">[4]</a></sup> Meanwhile, to accomplish the aim of time delay, which means we should make the GFP produce in different period of time, we decided to change the RBS&rsquo;s strength of <i>luxI</i> expression system. RBSes of different strengths will cause the speed of quorum sensing effect, which will lead to the delay of GFP production time.<sup><a href="#_ENREF_5" title="Ron Weiss, 2003 #2">[5]</a><a href="#_ENREF_6" title="Ron Weiss, 2003 #2">[6]</a></sup></p><br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p align="left">Based on our last year&rsquo;s project, for convenience of testing the results of time delay device, we chose GFP instead of CcdB as our reporter. On the other hand, because of the basal expression of <em>P<sub>lac</sub></em> is very strong, we decided to use arabinose to induce <em>P<sub>BAD</sub></em> promoter to activate our device.<sup><a href="#_ENREF_4" title="Ron Weiss, 2003 #2">[4]</a></sup> Meanwhile, to accomplish the aim of time delay, which means we should make the GFP produce in different period of time, we decided to change the RBS&rsquo;s strength of <i>luxI</i> expression system. RBSes of different strengths will cause the speed of quorum sensing effect, which will lead to the delay of GFP production time.<sup><a href="#_ENREF_5" title="Ron Weiss, 2003 #2">[5]</a><a href="#_ENREF_6" title="Ron Weiss, 2003 #2">[6]</a></sup></p><br /></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/<del class="diffchange diffchange-inline">3</del>/<del class="diffchange diffchange-inline">3c</del>/<del class="diffchange diffchange-inline">TimedelayFig_8</del>.<del class="diffchange diffchange-inline">png</del>" alt="" width="449" height="334" /><br /></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/<ins class="diffchange diffchange-inline">f</ins>/<ins class="diffchange diffchange-inline">fc</ins>/<ins class="diffchange diffchange-inline">Delay_02</ins>.<ins class="diffchange diffchange-inline">jpg</ins>" alt="" width="449" height="334" /><br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 6:</b> Fluorescence curves of the induced TD0.01, TD0.6, TD1.0 and <br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 6:</b> Fluorescence curves of the induced TD0.01, TD0.6, TD1.0 and <br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> the control BL21(DE3).<a target="_blank" href=" https://static.igem.org/mediawiki/2012/6/6f/-Fig_8-Comparison_of_TD0.01_0.6_1.0.jpg">[click to data]</a><br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> the control BL21(DE3).<a target="_blank" href=" https://static.igem.org/mediawiki/2012/6/6f/-Fig_8-Comparison_of_TD0.01_0.6_1.0.jpg">[click to data]</a><br /></div></td></tr>
</table>Youbinhttp://2012.igem.org/wiki/index.php?title=Team:XMU-China/timedelay&diff=269042&oldid=prevIcedcat at 06:01, 12 October 20122012-10-12T06:01:59Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="subtitle"><a name="_Toc02" id="Toc02"></a>2. Review of last year</span></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="subtitle"><a name="_Toc02" id="Toc02"></a>2. Review of last year</span></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p>Last year, we had developed a series of devices which program a bacteria population to maintain at different cell densities. <sup><a href="#_ENREF_2" title="Ron Weiss, 2003 #2">[2]</a></sup>We had designed and characterized the genetic circuit to establish a bacterial &lsquo;population-control&rsquo; device in <em>E. coli</em> based on the well-known quorum-sensing system from <em>Vibrio fischeri</em>, which autonomously regulates the density of an <i>E. coli</i> population. The cell density however was influenced by the expression levels of a killer gene (<i>ccdB</i>) in our device. As such, we had successfully controlled the expression levels of CcdB by using RBSes of different strength and mutated luxR promoters (<em>lux <del class="diffchange diffchange-inline">pr</del></em>). This circuit incorporated a mechanism for programmed death in respond to changes in the environment. <br /></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p>Last year, we had developed a series of devices which program a bacteria population to maintain at different cell densities. <sup><a href="#_ENREF_2" title="Ron Weiss, 2003 #2">[2]</a></sup>We had designed and characterized the genetic circuit to establish a bacterial &lsquo;population-control&rsquo; device in <em>E. coli</em> based on the well-known quorum-sensing system from <em>Vibrio fischeri</em>, which autonomously regulates the density of an <i>E. coli</i> population. The cell density however was influenced by the expression levels of a killer gene (<i>ccdB</i>) in our device. As such, we had successfully controlled the expression levels of CcdB by using RBSes of different strength and mutated luxR promoters (<em>lux <ins class="diffchange diffchange-inline">pR</ins></em>). This circuit incorporated a mechanism for programmed death in respond to changes in the environment. <br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </p><p align="center"> <img src="https://static.igem.org/mediawiki/2012/1/16/Timedelay_f1.png" alt="" width="508" height="349" /><br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </p><p align="center"> <img src="https://static.igem.org/mediawiki/2012/1/16/Timedelay_f1.png" alt="" width="508" height="349" /><br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 1:</b> The mechanism graph of iccdB last year<sup><a href="#_ENREF_3" title="Ron Weiss, 2003 #2">[3]</a></sup></p><hr><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 1:</b> The mechanism graph of iccdB last year<sup><a href="#_ENREF_3" title="Ron Weiss, 2003 #2">[3]</a></sup></p><hr><br></div></td></tr>
</table>Icedcathttp://2012.igem.org/wiki/index.php?title=Team:XMU-China/timedelay&diff=269041&oldid=prevIcedcat at 05:57, 12 October 20122012-10-12T05:57:39Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="subtitle"><a name="_Toc02" id="Toc02"></a>2. Review of last year</span></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="subtitle"><a name="_Toc02" id="Toc02"></a>2. Review of last year</span></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p>Last year, we had developed a series of devices which program a bacteria population to maintain at different cell densities. <sup><a href="#_ENREF_2" title="Ron Weiss, 2003 #2">[2]</a></sup>We had designed and characterized the genetic circuit to establish a bacterial &lsquo;population-control&rsquo; device in <em>E. coli</em> based on the well-known quorum-sensing system from <em>Vibrio fischeri</em>, which autonomously regulates the density of an <i>E. coli</i> population. The cell density however was influenced by the expression levels of a killer gene (<i>ccdB</i>) in our device. As such, we had successfully controlled the expression levels of <del class="diffchange diffchange-inline">ccdB </del>by using RBSes of different strength and mutated luxR promoters (<em>lux pr</em>). This circuit incorporated a mechanism for programmed death in respond to changes in the environment. <br /></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p>Last year, we had developed a series of devices which program a bacteria population to maintain at different cell densities. <sup><a href="#_ENREF_2" title="Ron Weiss, 2003 #2">[2]</a></sup>We had designed and characterized the genetic circuit to establish a bacterial &lsquo;population-control&rsquo; device in <em>E. coli</em> based on the well-known quorum-sensing system from <em>Vibrio fischeri</em>, which autonomously regulates the density of an <i>E. coli</i> population. The cell density however was influenced by the expression levels of a killer gene (<i>ccdB</i>) in our device. As such, we had successfully controlled the expression levels of <ins class="diffchange diffchange-inline">CcdB </ins>by using RBSes of different strength and mutated luxR promoters (<em>lux pr</em>). This circuit incorporated a mechanism for programmed death in respond to changes in the environment. <br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </p><p align="center"> <img src="https://static.igem.org/mediawiki/2012/1/16/Timedelay_f1.png" alt="" width="508" height="349" /><br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </p><p align="center"> <img src="https://static.igem.org/mediawiki/2012/1/16/Timedelay_f1.png" alt="" width="508" height="349" /><br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 1:</b> The mechanism graph of iccdB last year<sup><a href="#_ENREF_3" title="Ron Weiss, 2003 #2">[3]</a></sup></p><hr><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 1:</b> The mechanism graph of iccdB last year<sup><a href="#_ENREF_3" title="Ron Weiss, 2003 #2">[3]</a></sup></p><hr><br></div></td></tr>
</table>Icedcathttp://2012.igem.org/wiki/index.php?title=Team:XMU-China/timedelay&diff=269040&oldid=prevIcedcat at 05:54, 12 October 20122012-10-12T05:54:34Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/5/57/Timedelay_f2reversed.png" alt="" width="420" /><br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/5/57/Timedelay_f2reversed.png" alt="" width="420" /><br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 2:</b> The construction devices of time delay system<br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 2:</b> The construction devices of time delay system<br /></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p align="left">Based on our last year&rsquo;s project, for convenience of testing the results of time delay device, we chose GFP instead of <del class="diffchange diffchange-inline">ccdB </del>as our reporter. On the other hand, because of the basal expression of <em>P<sub>lac</sub></em> is very strong, we decided to use arabinose to induce <em>P<sub>BAD</sub></em> promoter to activate our device.<sup><a href="#_ENREF_4" title="Ron Weiss, 2003 #2">[4]</a></sup> Meanwhile, to accomplish the aim of time delay, which means we should make the GFP produce in different period of time, we decided to change the RBS&rsquo;s strength of <i>luxI</i> expression system. RBSes of different strengths will cause the speed of quorum sensing effect, which will lead to the delay of GFP production time.<sup><a href="#_ENREF_5" title="Ron Weiss, 2003 #2">[5]</a><a href="#_ENREF_6" title="Ron Weiss, 2003 #2">[6]</a></sup></p><br /></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p align="left">Based on our last year&rsquo;s project, for convenience of testing the results of time delay device, we chose GFP instead of <ins class="diffchange diffchange-inline">CcdB </ins>as our reporter. On the other hand, because of the basal expression of <em>P<sub>lac</sub></em> is very strong, we decided to use arabinose to induce <em>P<sub>BAD</sub></em> promoter to activate our device.<sup><a href="#_ENREF_4" title="Ron Weiss, 2003 #2">[4]</a></sup> Meanwhile, to accomplish the aim of time delay, which means we should make the GFP produce in different period of time, we decided to change the RBS&rsquo;s strength of <i>luxI</i> expression system. RBSes of different strengths will cause the speed of quorum sensing effect, which will lead to the delay of GFP production time.<sup><a href="#_ENREF_5" title="Ron Weiss, 2003 #2">[5]</a><a href="#_ENREF_6" title="Ron Weiss, 2003 #2">[6]</a></sup></p><br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/1/10/Timedelay_f3.png" alt="" width="428" height="241" /><br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/1/10/Timedelay_f3.png" alt="" width="428" height="241" /><br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 3:</b> The mechanism graph of time delay system</p><hr><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 3:</b> The mechanism graph of time delay system</p><hr><br></div></td></tr>
</table>Icedcathttp://2012.igem.org/wiki/index.php?title=Team:XMU-China/timedelay&diff=269039&oldid=prevIcedcat at 05:52, 12 October 20122012-10-12T05:52:48Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="subtitle"><a name="_Toc01" id="Toc01"></a>1. Overview</span></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="subtitle"><a name="_Toc01" id="Toc01"></a>1. Overview</span></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p>For the lengths of our genetic circuits are not the same, based on quorum sensing system <sup><a href="#_ENREF_1" title="Ron Weiss, 2003 #2">[1]</a></sup>we had studied last year, we considered that the time of genetic expressions of each circuit is different because of the different lengths and strength of ribosome binding sites. So we constructed circuits with <del class="diffchange diffchange-inline">five </del>grades strength of ribosome binding sites, including 0.01, 0.6 and 1.0. After induced by arabinose, the duration of response time for GFP expression increases as the strength of RBS declines, bringing about a time-course display.</p><hr></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p>For the lengths of our genetic circuits are not the same, based on quorum sensing system <sup><a href="#_ENREF_1" title="Ron Weiss, 2003 #2">[1]</a></sup>we had studied last year, we considered that the time of genetic expressions of each circuit is different because of the different lengths and strength of ribosome binding sites. So we constructed circuits with <ins class="diffchange diffchange-inline">three </ins>grades strength of ribosome binding sites, including 0.01, 0.6 and 1.0. After induced by arabinose, the duration of response time for GFP expression increases as the strength of RBS declines, bringing about a time-course display.</p><hr></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="subtitle"><a name="_Toc02" id="Toc02"></a>2. Review of last year</span></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="subtitle"><a name="_Toc02" id="Toc02"></a>2. Review of last year</span></div></td></tr>
</table>Icedcathttp://2012.igem.org/wiki/index.php?title=Team:XMU-China/timedelay&diff=229586&oldid=prevFBS at 01:18, 27 September 20122012-09-27T01:18:53Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 1:</b> The mechanism graph of iccdB last year<sup><a href="#_ENREF_3" title="Ron Weiss, 2003 #2">[3]</a></sup></p><hr><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 1:</b> The mechanism graph of iccdB last year<sup><a href="#_ENREF_3" title="Ron Weiss, 2003 #2">[3]</a></sup></p><hr><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="subtitle"><a name="_Toc03" id="Toc03"></a>3. Constructions this year</span></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="subtitle"><a name="_Toc03" id="Toc03"></a>3. Constructions this year</span></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/<del class="diffchange diffchange-inline">d</del>/<del class="diffchange diffchange-inline">dd</del>/<del class="diffchange diffchange-inline">Timedelay_f2</del>.png" alt="" width="<del class="diffchange diffchange-inline">399" height="285</del>" /><br /></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/<ins class="diffchange diffchange-inline">5</ins>/<ins class="diffchange diffchange-inline">57</ins>/<ins class="diffchange diffchange-inline">Timedelay_f2reversed</ins>.png" alt="" width="<ins class="diffchange diffchange-inline">420</ins>" /><br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 2:</b> The construction devices of time delay system<br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 2:</b> The construction devices of time delay system<br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p align="left">Based on our last year&rsquo;s project, for convenience of testing the results of time delay device, we chose GFP instead of ccdB as our reporter. On the other hand, because of the basal expression of <em>P<sub>lac</sub></em> is very strong, we decided to use arabinose to induce <em>P<sub>BAD</sub></em> promoter to activate our device.<sup><a href="#_ENREF_4" title="Ron Weiss, 2003 #2">[4]</a></sup> Meanwhile, to accomplish the aim of time delay, which means we should make the GFP produce in different period of time, we decided to change the RBS&rsquo;s strength of <i>luxI</i> expression system. RBSes of different strengths will cause the speed of quorum sensing effect, which will lead to the delay of GFP production time.<sup><a href="#_ENREF_5" title="Ron Weiss, 2003 #2">[5]</a><a href="#_ENREF_6" title="Ron Weiss, 2003 #2">[6]</a></sup></p><br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p align="left">Based on our last year&rsquo;s project, for convenience of testing the results of time delay device, we chose GFP instead of ccdB as our reporter. On the other hand, because of the basal expression of <em>P<sub>lac</sub></em> is very strong, we decided to use arabinose to induce <em>P<sub>BAD</sub></em> promoter to activate our device.<sup><a href="#_ENREF_4" title="Ron Weiss, 2003 #2">[4]</a></sup> Meanwhile, to accomplish the aim of time delay, which means we should make the GFP produce in different period of time, we decided to change the RBS&rsquo;s strength of <i>luxI</i> expression system. RBSes of different strengths will cause the speed of quorum sensing effect, which will lead to the delay of GFP production time.<sup><a href="#_ENREF_5" title="Ron Weiss, 2003 #2">[5]</a><a href="#_ENREF_6" title="Ron Weiss, 2003 #2">[6]</a></sup></p><br /></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p><span class="subsubtitle">4.1 Optimization of inducer concentration</span><br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p><span class="subsubtitle">4.1 Optimization of inducer concentration</span><br /></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del><p align="center"> <img src="https://static.igem.org/mediawiki/2012/0/0f/TimedelayFig4.png" alt="" width="491" height="347" /><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins><p align="center" > <img src="https://static.igem.org/mediawiki/2012/0/0f/TimedelayFig4.png" alt="" width="491" height="347" /><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del><b>Figure 4:</b> The optimization of the arabinose concentration added into the TD1.0 including 0, 0.1, 1.0, 10 mM arabinose.<a target="_blank" href=" https://static.igem.org/mediawiki/2012/e/e4/-Fig_4-Optimization_of_arabinose_induced_concentration_%28TD1.0%29.jpg">[click to data]</a></p><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><b>Figure 4:</b> The optimization of the arabinose concentration added into the TD1.0 including<ins class="diffchange diffchange-inline"><br></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>0, 0.1, 1.0, 10 mM arabinose.<a target="_blank" href=" https://static.igem.org/mediawiki/2012/e/e4/-Fig_4-Optimization_of_arabinose_induced_concentration_%28TD1.0%29.jpg">[click to data]</a></p><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p align="left">We can find that the curve with 0.1mM arabinose has a higher fluorescence. So we chose the </a>0.1 mM arabinose for the following fluorescence tests.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p align="left">We can find that the curve with 0.1mM arabinose has a higher fluorescence. So we chose the </a>0.1 mM arabinose for the following fluorescence tests.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p class="subsubtitle">4.2 Fluorescent test of the time delay devices</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p class="subsubtitle">4.2 Fluorescent test of the time delay devices</p></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/8/8c/XMUtimedelay5_6_7.png" alt="" width="<del class="diffchange diffchange-inline">490</del>" /><br /></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/8/8c/XMUtimedelay5_6_7.png" alt="" width="<ins class="diffchange diffchange-inline">500</ins>" /><br /></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <b>Figure 5:</b> Fluorescence curves of strain TD1.0, TD0.6 and TD0.01 induced with 0.1 mM arabinose and without arabinose.<a target="_blank" href=" https://static.igem.org/mediawiki/2012/6/6a/XMUtimedelay5_6_7data.png">[click to data]</a><br /></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <b>Figure 5:</b> Fluorescence curves of strain TD1.0, TD0.6 and TD0.01 <ins class="diffchange diffchange-inline"><br></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins>induced</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins>with 0.1 mM arabinose and without arabinose.<a target="_blank" href=" https://static.igem.org/mediawiki/2012/6/6a/XMUtimedelay5_6_7data.png">[click to data]</a><br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </p></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/3/3c/TimedelayFig_8.png" alt="" width="449" height="334" /><br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p align="center"><img src="https://static.igem.org/mediawiki/2012/3/3c/TimedelayFig_8.png" alt="" width="449" height="334" /><br /></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <b>Figure 6:</b> Fluorescence curves of the induced TD0.01, TD0.6, TD1.0 and the control BL21(DE3).<a target="_blank" href=" https://static.igem.org/mediawiki/2012/6/6f/-Fig_8-Comparison_of_TD0.01_0.6_1.0.jpg">[click to data]</a><br /></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <b>Figure 6:</b> Fluorescence curves of the induced TD0.01, TD0.6, TD1.0 and <ins class="diffchange diffchange-inline"><br></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins>the control BL21(DE3).<a target="_blank" href=" https://static.igem.org/mediawiki/2012/6/6f/-Fig_8-Comparison_of_TD0.01_0.6_1.0.jpg">[click to data]</a><br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2012/0/00/TimedelayFig_9.png" alt="" width="480" height="371" /><br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2012/0/00/TimedelayFig_9.png" alt="" width="480" height="371" /><br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 7:</b>The separate time taken up for the three circuits' fluorescent value to reach 5000 <br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <b>Figure 7:</b>The separate time taken up for the three circuits' fluorescent value to reach 5000 <br></div></td></tr>
</table>FBShttp://2012.igem.org/wiki/index.php?title=Team:XMU-China/timedelay&diff=213342&oldid=prevYoubin at 19:56, 26 September 20122012-09-26T19:56:15Z<p></p>
<a href="http://2012.igem.org/wiki/index.php?title=Team:XMU-China/timedelay&diff=213342&oldid=212210">Show changes</a>Youbinhttp://2012.igem.org/wiki/index.php?title=Team:XMU-China/timedelay&diff=212210&oldid=prevYoubin at 19:32, 26 September 20122012-09-26T19:32:09Z<p></p>
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</table>Youbinhttp://2012.igem.org/wiki/index.php?title=Team:XMU-China/timedelay&diff=197093&oldid=prevIcedcat at 13:23, 26 September 20122012-09-26T13:23:47Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Figure 6. Fluorescence curves of the induced strain TD0.01, TD0.6, TD1.0 and <em>E. coli</em> BL21(DE3)<br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Figure 6. Fluorescence curves of the induced strain TD0.01, TD0.6, TD1.0 and <em>E. coli</em> BL21(DE3)<br /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2012/0/00/TimedelayFig_9.png" alt="" width="480" height="371" /><br /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2012/0/00/TimedelayFig_9.png" alt="" width="480" height="371" /><br /></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Figure 7. <del class="diffchange diffchange-inline">Experimentally measured the average </del>time <del class="diffchange diffchange-inline">of </del>the three <del class="diffchange diffchange-inline">cultures&rsquo; fluorescence up </del>to 5000 <del class="diffchange diffchange-inline"><br></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> Figure 7. <ins class="diffchange diffchange-inline">The separate </ins>time <ins class="diffchange diffchange-inline">taken up for </ins>the three <ins class="diffchange diffchange-inline">circuits' fluorescent value </ins>to <ins class="diffchange diffchange-inline">reach </ins>5000 </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del>after induction, <del class="diffchange diffchange-inline">it </del>shows that the stronger RBS circuit contains, <del class="diffchange diffchange-inline"> </del>the less time it takes.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>after induction, <ins class="diffchange diffchange-inline">which </ins>shows that the stronger RBS circuit contains, the less time it takes.</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p>Compared with the groups absence of arabinose, the fluorescence of <del class="diffchange diffchange-inline">induction </del>groups were much higher. On the other hand, we could find even the circuit with RBS<sub>0.01</sub>, the most insensitive one, also expressed green fluorescence protein without arabinose, which means basal expressing phenomenon. According to figure 5, when the three cultures induced by the same concentration of arabinose, the fluorescence of strain TD1.0 was the highest, strain TD0.6 was middle and strain TD0.01 was the lowest. Finally, as shown in figure 8, the fluorescence of the circuits with different RBSes also reached a certain level in different time. It fit the goal of time delay.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p>Compared with the groups absence of arabinose, the fluorescence of <ins class="diffchange diffchange-inline">inducted </ins>groups were much higher. On the other hand, we could find even the circuit with RBS<sub>0.01</sub>, the most insensitive one, also expressed green fluorescence protein without arabinose, which means basal expressing phenomenon. According to figure 5, when the three cultures induced by the same concentration of arabinose, the fluorescence of strain TD1.0 was the highest, strain TD0.6 was middle and strain TD0.01 was the lowest. Finally, as shown in figure 8, the fluorescence of the circuits with different RBSes also reached a certain level in different time. It fit the goal of time delay.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div><hr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div><hr></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p class="subtitle"><a name="_Toc05" id="Toc05"></a>5. Reference</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p class="subtitle"><a name="_Toc05" id="Toc05"></a>5. Reference</div></td></tr>
</table>Icedcat