Team:UT-Tokyo/CTCD/Content

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<li><a href="Javascript:loadContent('Modeling-block','Modeling-1')">BBa_K747096</a></li>
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<li><a href="#">Contents1</a></li>
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<img src ="https://static.igem.org/mediawiki/2014/1/1a/SubCTC_BBaK747096.png" class = "contTitle" />
 
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<h3>Characterization of BBa_K747096</h3>
 
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<p>During DNA assembly, we found that this promoter was activated in Escherichia coli. Then we decided to characterize this promoter activity by comparing with constitutive promoter (BBa_J23101) using GFP (BBa_E0040) (Figure 1).<br>As seen in Figure 1, this promoter shows weaker activity than BBa_J23101.</p>
 
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<h3>Figure 1</h3>
 
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<p>The activity of BBa_K747096GFP was activated by 501nm excitation laser. <br>a)Measured when OD600 is 0.9~1.1.<br>b)Measured after cultured O/N.</p>
 
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<p>In order to calculate RPU (relative promoter unit [1]), we performed real-time measurement of GFP fluorescence (Figure 2). We measured activity of GFP and OD600, which are necessary for calculation of RPU.</p>
 
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<h3>Figure 2</h3>
 
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<p>Real-time measurement of BBa_K747096<br>a)Activity of GFP (activated by 488nm excitation laser).<br>b)OD600. </p>
 
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<p>We calculated RPU(Relative Promoter Unit) of BBa_K747096 from the Fig. 2 using the equation (1).</p>
 
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<img src="https://static.igem.org/mediawiki/2014/d/d1/Cocoa_equation_1.png" class="math" height="60px" />
 
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<p>This equation can be used only when Fluorescence per cell is in steady state. In this experiment, this condition was fulfilled as shown in Table1.</p>
 
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<table border="1" width="850" align="center"> <caption>Table 1 : Fluorescence / OD</caption>
 
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<tbody>
 
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<tr bgcolor="#cccccc">
 
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<th></th><th>K747096</th><th>J23101</th></tr>
 
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<tr align="center"><td>t=4h</td><td>92.62</td><td>13.14</td></tr>
 
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<tr align="center"><td>t=6h</td><td>99.37</td><td>19.86</td></tr>
 
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</tbody>
 
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</table>
 
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<p>From Table1, we can get RPU of BBa_K747096:</p>
 
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<p>RPU = 5.8</p>
 
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<p>[1]:https://2010.igem.org/Team:Kyoto/LearnMore#Relative_Promoter_Unit_.28RPU.29</p>
 
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Revision as of 06:38, 16 October 2014