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| - | <a href="https://2013.igem.org/Team:Tianjin/Project">Overview</a>
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| - | <a href="https://2013.igem.org/Team:Tianjin/Project/Background">Background</a>
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| - | <a href="https://2013.igem.org/Team:Tianjin/Project/Experiment/Endo-Induce">Experiment&Result</a>
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| - | <a href="https://2013.igem.org/Team:Tianjin/Project/Experiment/Endo-Induce">In vivo alkane sensing test</a>
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| - | </li>
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| - | <a href="https://2013.igem.org/Team:Tianjin/Project/Experiment/Exo-Induce">Exogenous Alkanes Induce</a>
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| - | <a href="https://2013.igem.org/Team:Tianjin/Project/Experiment/TetA">Endogenous Induce&Resistance Selection</a>
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| - | <a href="https://2013.igem.org/Team:Tianjin/Project/Experiment/LibraryConstruction">Library Construction</a>
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| - | <center><span style="font-family:Arial;font-size:46px;color:#000;"> Experiment</span></center>
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| - | = Endogenous Alkanes Induce=
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| - | <div style="text-align:center;vertical-align:middle;"><a href="https://static.igem.org/mediawiki/2013/c/c3/Tju-endo1.jpg" target="_blank" ><img src="https://static.igem.org/mediawiki/2013/c/c3/Tju-endo1.jpg" width="400px" /></a></div>
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| - | <p> After systematic analysis of the microbial synthesis pathway from glucose to alkane in E.coli, we decide to choose the route from Fatty Acyl-ACP to alkane as the alkane producing module. Meanwhile, we conduct codon-optimization to the genes encoding NPDC and AAR based on the work done by Andreas Schirmer, making them more applicable to E.coli.</p>
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| - | <p> Here, we want to test the response of Alk-Sensor to endogenous alkanes.
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| - | Construct:</p>
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| - | <p> We constructed 3 strains as shown in the figure below.</p>
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| - | <br/>
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| - | <div style="text-align:center;vertical-align:middle;"><a href="https://static.igem.org/mediawiki/2013/d/d4/Tju-endo2.jpg" target="_blank" ><img src="https://static.igem.org/mediawiki/2013/d/d4/Tju-endo2.jpg" width="700px" /></a></div>
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| - | <p>strain1:No.53：producing module (constitutive promoter J23100)+ Alk-Sensor device</p>
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| - | <p>strain2:No.54：producing module (constitutive promoter J23114) + Alk-Sensor device</p>
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| - | <p>strain3:No.55：Null+ Alk-Sensor device</p>
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| - | <p> The promoter strength of No.54 is stronger than that of No.53. Theorectically, alkane productivity of No.53 and No.54 will be different, and intracellular alkane concentration of the above 3 strains will be different-- No.54 the highest, No.53 lower and No.55 the lowest.</p>
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| - | <p> The 3 strains are cultured in 3ml M9 medium for about 24 hours.</p>
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| - | <div style="text-align:center;vertical-align:middle;"><a href="https://static.igem.org/mediawiki/2013/0/08/Tju-Endo3.png" target="_blank" ><img src="https://static.igem.org/mediawiki/2013/0/08/Tju-Endo3.png" width="706px" /></a></div>
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| - | <div><center><b>Figure 1.</b> Cell Induced-Fluorescent Indensity </center></div>
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| - | <p> <b>Result：</b>As shown in the diagram above, the fluorescent indensity of No.54 is the strongest, No.53 weaker and No.55 the weakest.</p>
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| - | <p ><b>Conclusion:</b></p>
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| - | <p>(1) Alk-Sensor device can respond to the mixed intracellular alkanes, which can be used to detect alkanes inside the cell.</p>
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| - | <p>(2) There is a positive correlation between the concentration of intracellular alkanes, and the expression level of the reporter.</p>
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| - | E-mail：austinamens@gmail.com | Address:Building No.20, No.92 Weijin Road, Tianjin, China | Zip-code：300072
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| - | <br />Copyright 2013 © Tianjin University iGEM Team
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