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+ | <div id="HOME"> | ||
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+ | <ul><li><a href="https://2017.igem.org/Team:Nanjing-China/Demonstrate">Results</a></ul></li></div> | ||
+ | <ul> | ||
+ | <li><a href="#ch2o">CH<sub>2</sub>O</a></li> | ||
+ | <li><a href="#h2s">H<sub>2</sub>S</a></li> | ||
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− | < | + | <li><a href="https://2017.igem.org/Team:Nanjing-China">HOME</a> |
− | < | + | <ul> |
− | <ul> | + | <li><a href="https://2017.igem.org/Team:Nanjing-China">Introduction</a></li> |
− | <li><a href="https:// | + | <li><a href="https://2017.igem.org/Team:Nanjing-China/Background">Background</a></li> |
− | <li><a href="https:// | + | </ul> |
− | <li><a href="https:// | + | </li> |
− | </ul> | + | <li><a href="https://2017.igem.org/Team:Nanjing-China/Parts">PARTS</a> |
− | + | <li><a href="https://2017.igem.org/Team:Nanjing-China/Sliver">JUDGE</a> | |
+ | <ul> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/Sliver">Sliver</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/Gold">Gold</a></li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/Design">PROJECT</a> | ||
+ | <ul> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/Design">Design</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/Notebook">Notebook</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/Results">Results</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/Demonstrate">Demonstrate</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/Improvement">Improvement</a></li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/TEAM/Introduction">TEAM</a> | ||
+ | <ul> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/TEAM/Introduction">Introduction</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/Members">Members</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/Attributions">Attributions</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/Collaborations">Collaboration</a></li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/HP/Silver">OTHERS</a> | ||
+ | <ul> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/HP/Silver">HP-Silver</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/HP/Gold_Integrated">HP-Gold</a></li> | ||
+ | <li><a href="https://2017.igem.org/Team:Nanjing-China/Model">Model</a></li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="contain" onmousemove="MM_changeProp('d','','left','85%','DIV')"> | ||
+ | <div class="word" style=" position:relative; top:-80px;"> | ||
+ | <h1>Results</h1> | ||
+ | <p style="font-size:103%;">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. </p> | ||
+ | </div> | ||
+ | <div id="ch2o"> | ||
+ | <div align="center"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/f/f1/T-Nanjing-China-project-ch2o.png" width="35%" /> | ||
+ | <table width="80%" border="0" cellspacing="1" cellpadding="1"> | ||
+ | <tr> | ||
+ | <td colspan="2"><blockquote> | ||
+ | <p>We have designed a formaldehyde sensor sequence, which is a part of our team .</p> | ||
+ | <p>The sequence is composed of PfrmR, gene frmR, flag tag, PfrmAB, gene RFP.</p> | ||
+ | <p>When the pathway works, we can see that the E.coli turns to red with naked eyes at the presence of formaldehyde. </p> | ||
+ | </blockquote></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td colspan="2"><div align="center"><img src="https://static.igem.org/mediawiki/2017/b/b4/T-Nanjing-China-ch2o-2.png" width="500" height="75" /></div></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td colspan="2"><blockquote> | ||
+ | <p>In order to demonstrate the design, a lot of experiments have been done.</p></blockquote></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><div align="center"><p><font size="-1">Figure 3. Influence of Formaldehyde Induce Time on Fluorescence Expression</font></p><img src="https://static.igem.org/mediawiki/2017/8/88/T-Nanjing-China-ch2o-8.png" width="350" height="251" /></div></td> | ||
+ | <td><div align="center"> | ||
+ | <p><img src="https://static.igem.org/mediawiki/2017/0/04/T-Nanjing-China-ch2o-9.png" width="350" height="245" /></p> | ||
+ | <p><font size="-1">Figure4.A photograph of E.coli cells containing the formaldehyde-induced RFP expression plasmid, or without formaldehyde induction, and re-suspended in PBS buffer(pH7.4)</font></p> | ||
+ | </div></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><div align="center"><img src="https://static.igem.org/mediawiki/2017/7/76/T-Nanjing-China-ch2o-10.png" width="350" height="265" /></div></td> | ||
+ | <td><div align="center"><img src="https://static.igem.org/mediawiki/2017/9/98/T-Nanjing-China-ch2o-11.png" width="350" height="265" /></div></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td colspan="2"><blockquote> | ||
+ | <p><font size="-1">Figure5.Fluorescence measurement of E.coli cells containing the formaldehyde-induced RFP expression plasmid after gradient concentrations of formaldehyde induction and re-suspended in PBS buffer(pH7.4)</font></p> | ||
+ | </blockquote></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td colspan="2">It is worth to be mentioned that the team OUC help us demonstrate the result.</td></tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div id="h2s"> | ||
+ | <div align="center"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/c/c7/T-Nanjing-China-project-h2s.png" width="40%"/> | ||
+ | <table width="80%" border="0" cellspacing="1" cellpadding="1"> | ||
+ | <tr> | ||
+ | <td colspan="2"><blockquote><p>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. </p> | ||
+ | <p>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.</p> | ||
+ | </blockquote></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td colspan="2"><div align="center"><img src="https://static.igem.org/mediawiki/2017/d/d8/T-Nanjing-China-h2s-2.png" width="600" height="90" /></div></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td colspan="2">In the experiment, we proved that the sequence worked well and was useful to detect hydrogen sulfide</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><div align="center"><img src="https://static.igem.org/mediawiki/2017/3/3c/T-Nanjing-China-h2s-4-1.jpg" width="400" /></div></td> | ||
+ | <td><div align="center"><img src="https://static.igem.org/mediawiki/2017/c/c6/T-Nanjing-China-h2s-5.png" width="400" /> | ||
+ | <p><font size="-1">Figure1.Whole-cell sequence dual-enzyme digestion</font></p></div></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><p><font size="-1">a)</font></p><img src="https://static.igem.org/mediawiki/2017/6/63/T-Nanjing-China-h2s-6.png" width="400" /> | ||
+ | </td> | ||
+ | <td><p><font size="-1">b)</font></p><div align="center"><img src="https://static.igem.org/mediawiki/2017/4/4d/T-Nanjing-China-h2s-8.png" width="400" /><p><font size="-1"></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><p><font size="-1">c)</font></p><img src="https://static.igem.org/mediawiki/2017/b/bc/T-Nanjing-China-h2s-9.png" width="400" /><p><font size="-1"></p></p></td> | ||
+ | <td><p><font size="-1">a)RFP responsiveness of the detector system.<br /> | ||
+ | b) A visible photograph of a).<br /> | ||
+ | c) Test of selectivity.</font></p></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div id="h2"> | ||
+ | <div align="center"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/0/09/T-Nanjing-China-project-h2.png" width="30%"/> | ||
+ | <table width="80%" border="0" cellspacing="1" cellpadding="1"> | ||
+ | <tr> | ||
+ | <td colspan="2"> | ||
+ | <p>There is a composite of hydrogen sensor full length sequence. </p> | ||
+ | <p>The order of the elements is: HoxA-HoxB-HoxC-HoxJ-terminator-HoxP-EGFP. The sequence of HoxABCJP comes from Ralstonia eutropha H16 megaplasmid pHG1. The hole sequence acts as an hydrogen sensor.</p> | ||
+ | <p>When the amount of hydrogen goes to a higher level, Fluorescence intensity increases apparently.</p> | ||
+ | </td></tr> | ||
+ | <tr> | ||
+ | <td colspan="2"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/b/b5/T-Nanjing-China-h2-2.png" width="650" height="100" /> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td colspan="2"> | ||
+ | <p>The sequence was a good detecter in the lab work.</p> | ||
+ | </td></tr> | ||
+ | <tr> | ||
+ | <td><div align="center"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/1/10/T-Nanjing-China-h2-7.png" width="350" height="333" /> | ||
+ | <p><font size="-1">Figure1. Coomassie Brilliant Blue R-250-stained SDS-Page analysis of recombinant E.coli expressing hoxABCJ-terminator-hoxp-gfp</font></p> | ||
+ | </div></td> | ||
+ | <td><div align="center"><img src="https://static.igem.org/mediawiki/2017/9/9f/T-Nanjing-China-h2-8.png" width="350" height="411" /> | ||
+ | <p><font size="-1">Fingure 2. Western blot analysis of recombinant E.coli expressing his-hoxA</font></p></div></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td colspan="2"><blockquote> | ||
+ | <p>Fluorescence intensity remains stationary when IPTG is added.<br /> | ||
+ | And Fluorescence intensity increases in a low hydrogen atmosphere.<br /> | ||
+ | When the amount of hydrogen goes to a higher level Fluorescence intensity increases apparently. meaning the designed reporter pathway have worked. </p> | ||
+ | </blockquote></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td colspan="2"><div align="center"><img src="https://static.igem.org/mediawiki/2017/6/69/T-Nanjing-China-h2-9.png" width="450" height="250" /> | ||
+ | <p><font size="-1">Figure 3. Influence of H2 concentration on fluorescence expression</font></p> | ||
+ | </div></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="footer"> | ||
+ | <div align="center"> | ||
+ | <div class="f"> | ||
+ | <div class="f-word"><p><strong><u>Address</u></strong></p> | ||
+ | <p>Life Science Department<br /> | ||
+ | #163 Xianlin Blvd, Qixia District<br /> | ||
+ | Nanjing University<br /> | ||
+ | Nanjing, Jiangsu Province<br /> | ||
+ | P.R. of China<br /> | ||
+ | Zip: 210046 | ||
+ | </p> | ||
+ | <p><strong><u>Email</u></strong></p> | ||
+ | <p>nanjing_china@163.com</p> | ||
+ | <p><strong><u>Social media</u></strong></p> | ||
+ | <p>Twitter button: <br/> | ||
+ | <a href="https://twitter.com/Guja1501">iGEM Nanjing-China@nanjing_china</a></p> | ||
+ | </div> | ||
+ | <div class="f-img"> | ||
+ | <div align="center"><img src="https://static.igem.org/mediawiki/2017/0/04/T-Nanjing-China-%E5%8D%97%E4%BA%AC%E5%A4%A7%E5%AD%A6.png" name="img1" width="160" height="220" id="img1" onMouseOver="MM_changeProp('img1','','width','180px','IMG');MM_changeProp('img1','','height','240px','IMG')" onMouseOut="MM_changeProp('img1','','width','160px','IMG');MM_changeProp('img1','','height','220px','IMG')" /></div> | ||
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+ | </div> | ||
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+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="f-b"><img src="https://static.igem.org/mediawiki/2017/9/91/T-Nanjing-China-foot.png" width="100%"/></div> | ||
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Revision as of 04:21, 28 October 2017
Results
In the part of lab work, we have designed three biosensor sequence and improved an old part, J23000. What's more, all the three design have been demonstrate by us.
|
|
|
|
Figure 3. Influence of Formaldehyde Induce Time on Fluorescence Expression |
Figure4.A photograph of E.coli cells containing the formaldehyde-induced RFP expression plasmid, or without formaldehyde induction, and re-suspended in PBS buffer(pH7.4) |
|
|
It is worth to be mentioned that the team OUC help us demonstrate the result. |
|
|
In the experiment, we proved that the sequence worked well and was useful to detect hydrogen sulfide | |
Figure1.Whole-cell sequence dual-enzyme digestion |
|
a) |
b) |
c) |
a)RFP responsiveness of the detector system. |
There is a composite of hydrogen sensor full length sequence. The order of the elements is: HoxA-HoxB-HoxC-HoxJ-terminator-HoxP-EGFP. The sequence of HoxABCJP comes from Ralstonia eutropha H16 megaplasmid pHG1. The hole sequence acts as an hydrogen sensor. When the amount of hydrogen goes to a higher level, Fluorescence intensity increases apparently. | |
The sequence was a good detecter in the lab work. | |
Figure1. Coomassie Brilliant Blue R-250-stained SDS-Page analysis of recombinant E.coli expressing hoxABCJ-terminator-hoxp-gfp |
Fingure 2. Western blot analysis of recombinant E.coli expressing his-hoxA |
|
|
Figure 3. Influence of H2 concentration on fluorescence expression |