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- <a href="https://2017.igem.org/Team:Nanjing-China/Design">Design</a>
- <a href="https://2017.igem.org/Team:Nanjing-China/Notebook">Notebook</a>
- <a href="https://2017.igem.org/Team:Nanjing-China/Demonstrate">Results</a>
- <a href="#ch2o">CH2O</a>
- <a href="#h2s">H2S</a>
- <a href="#h2">H2</a>
- <a href="https://2017.igem.org/Team:Nanjing-China/Demonstrate">Demonstrate</a>
- <a href="https://2017.igem.org/Team:Nanjing-China/Improvement">Improvement</a>
In the part of lab work, we have <a href="https://2017.igem.org/Team:Nanjing-China/Design">designed</a> three biosensor <a href="https://2017.igem.org/Team:Nanjing-China/Parts">sequence</a> and <a href="https://2017.igem.org/Team:Nanjing-China/PP">improved</a> an old part, <a href="http://parts.igem.org/Part:BBa_J23000">J23000</a>. What's more, all the three design have been <a href="https://2017.igem.org/Team:Nanjing-China/Demonstrate">demonstrate</a> by us.
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We have designed a formaldehyde sensor sequence, which is a part of our team . The sequence is composed of PfrmR, gene frmR, flag tag, PfrmAB, gene RFP. When the pathway works, we can see that the E.coli turns to red with naked eyes at the presence of formaldehyde. |
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<img src="" width="400" height="286" /> Figure 3. Influence of Formaldehyde Induce Time on Fluorescence Expression |
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Figure6. Optical density(600nm) of (a) Escherichia coli BL21 and (b) recombinant bioluminescent Escherichia coli BL21 harboring frmR-RFP fusion after 10 hours’ incubation with 800uM different aldehydes |
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Figure8.Fluorescence test of various aldehydes using recombinant bioluminescent Escherichia coli BL21 |
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Figure9.The tolerance of recombinant bioluminescent Escherichia coli BL21 to various concentration of formaldehyde |
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It is worth to be mentioned that the team OUC help us demonstrate the result. |
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As to the hydrogen sulfide sensor, we also designed a whole-cell biocatalytic system, displaying the concentration of hydrogen sulfide by the compound’s influence on specific genes’ expression in modified E.coli. In our design, we use red fluorescent protein as the indicator.When hydrogen sulfide exits, the gene transcription is activated, and the bacteria turns red. |
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In the experiment, we proved that the sequence worked well and was useful to detect hydrogen sulfide | |||||||||||||
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Figure1.Whole-cell sequence dual-enzyme digestion |
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a) <img src="" width="400" /> |
b) <img src="" width="400" /> </td>
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<p>c)</p><img src="" width="400" /><p></p></p></td> | <p>Figure 2.a)RFP responsiveness of the detector system.b) A visible photograph of a).</p></td> </tr> </table> </div> </div> <img src="" width="30%"/>
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