Difference between revisions of "Team:Nanjing-China/Results"

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<title>Team:Nanjing-China - 2017.igem.org</title>
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<h1>Results</h1>
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<p>Here you can describe the results of your project and your future plans. </p>
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<h5>What should this page contain?</h5>
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<ul>
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<li> Clearly and objectively describe the results of your work.</li>
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<li> Future plans for the project. </li>
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<li> Considerations for replicating the experiments. </li>
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<h5>You should also describe what your results mean: </h5>
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<script type="text/javascript" src="https://2017.igem.org/Template:Nanjing-China/Javascript/d6?action=raw&ctype=text/javascript"></script>
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<li> Interpretation of the results obtained during your project. Don't just show a plot/figure/graph/other, tell us what you think the data means. This is an important part of your project that the judges will look for. </li>
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<li> Show data, but remember all measurement and characterization data must be on part pages in the Registry. </li>
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<li> Consider including an analysis summary section to discuss what your results mean. Judges like to read what you think your data means, beyond all the data you have acquired during your project. </li>
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<div id="Layer1" style="visibility: hidden"><img src="https://static.igem.org/mediawiki/2017/4/4e/T-Nanjing-China-cb4.png" width="220" /></div>
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<div id="Layer2" style="left: 79px; top: 83px; visibility: hidden;"><img src="https://static.igem.org/mediawiki/2017/1/16/T-Nanjing-China-star.png" width="40" /></div>
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<div align="center">
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<div id="HOME">
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<div class="sub">
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  <ul><li><a href="https://2017.igem.org/Team:Nanjing-China/Demonstrate">Results</a></ul></li></div>
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    <ul>
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    <li><a href="#ch2o">CH<sub>2</sub>O</a></li>
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  <li><a href="#h2s">H<sub>2</sub>S</a></li>
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    <li><a href="#h2">H<sub>2</sub></a></li>
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  </ul>
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<h5> Project Achievements </h5>
 
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<p>You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.</p>
 
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<ul>
 
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<li>A list of linked bullet points of the successful results during your project</li>
 
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<li>A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.</li>
 
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    <ul>  
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<h5>Inspiration</h5>
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  <li><a href="https://2017.igem.org/Team:Nanjing-China">HOME</a>
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<p>See how other teams presented their results.</p>
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    <ul>
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<ul>
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        <li><a href="https://2017.igem.org/Team:Nanjing-China">Introduction</a></li>
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<li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results/Pathway">2014 TU Darmstadt </a></li>
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            <li><a href="https://2017.igem.org/Team:Nanjing-China/Background">Background</a></li>
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<li><a href="https://2014.igem.org/Team:Imperial/Results">2014 Imperial </a></li>
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</ul>
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<li><a href="https://2014.igem.org/Team:Paris_Bettencourt/Results">2014 Paris Bettencourt </a></li>
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    </li>
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</ul>
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    <li><a href="https://2017.igem.org/Team:Nanjing-China/Parts">PARTS</a>
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    <li><a href="https://2017.igem.org/Team:Nanjing-China/Sliver">JUDGE</a>
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    <ul>
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        <li><a href="https://2017.igem.org/Team:Nanjing-China/Sliver">Sliver</a></li>
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            <li><a href="https://2017.igem.org/Team:Nanjing-China/Gold">Gold</a></li>
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</ul>
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      </li>
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    <li><a href="https://2017.igem.org/Team:Nanjing-China/Design">PROJECT</a>
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    <ul>
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        <li><a href="https://2017.igem.org/Team:Nanjing-China/Design">Design</a></li>
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                <li><a href="https://2017.igem.org/Team:Nanjing-China/Notebook">Notebook</a></li>
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                <li><a href="https://2017.igem.org/Team:Nanjing-China/Results">Results</a></li>
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                <li><a href="https://2017.igem.org/Team:Nanjing-China/Demonstrate">Demonstrate</a></li>
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            <li><a href="https://2017.igem.org/Team:Nanjing-China/Improvement">Improvement</a></li>
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</ul>
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        </li>
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        <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>
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                <li><a href="https://2017.igem.org/Team:Nanjing-China/Attributions">Attributions</a></li>
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                <li><a href="https://2017.igem.org/Team:Nanjing-China/Collaborations">Collaboration</a></li>
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</ul>
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    </li>
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    <li><a href="https://2017.igem.org/Team:Nanjing-China/HP/Silver">OTHERS</a>
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    <ul>
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        <li><a href="https://2017.igem.org/Team:Nanjing-China/HP/Silver">HP-Silver</a></li>
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                <li><a href="https://2017.igem.org/Team:Nanjing-China/HP/Gold_Integrated">HP-Gold</a></li>
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            <li><a href="https://2017.igem.org/Team:Nanjing-China/Model">Model</a></li>
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</ul>
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    </li>
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      </ul>
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<div class="contain"  onmousemove="MM_changeProp('d','','left','85%','DIV')">
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<div class="word" style=" position:relative; top:-80px;">
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    <h1>Results</h1>
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    <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>
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    </div>
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      <div id="ch2o">
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        <div align="center">
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            <img src="https://static.igem.org/mediawiki/2017/f/f1/T-Nanjing-China-project-ch2o.png" width="35%" />
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            <table width="80%" border="0" cellspacing="1" cellpadding="1">
 +
            <tr>
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              <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>
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            </tr>
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            <tr>
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              <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>
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              </tr>
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            <tr>
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            <td colspan="2"><blockquote>
 +
              <p>In order to demonstrate the design, a lot of experiments have been done.</p></blockquote></td>
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            </tr>
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            <tr>
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              <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>
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              <td><div align="center">
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                <p><img src="https://static.igem.org/mediawiki/2017/0/04/T-Nanjing-China-ch2o-9.png" width="350" height="245" /></p>
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              <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>
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              </div></td>
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            </tr>
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            <tr>
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            </tr>
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            <tr>
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              <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>
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              <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>
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            </tr>
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            <tr>
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            <td colspan="2"><blockquote>
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              <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>
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          </tr>
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          <tr>
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            <td colspan="2">It is worth to be mentioned that the team OUC help us demonstrate the result.</td></tr>
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          </table>
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        </div>
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      </div>
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        <div id="h2s">
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        <div align="center">
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          <img src="https://static.igem.org/mediawiki/2017/c/c7/T-Nanjing-China-project-h2s.png" width="40%"/>
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          <table width="80%" border="0" cellspacing="1" cellpadding="1">
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            <tr>
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            <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&rsquo;s influence on specific genes&rsquo; 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>
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            <tr>
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              <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>
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            </tr>
 +
              <tr>
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              <td colspan="2">In the experiment, we proved that the sequence worked well and was useful to detect hydrogen sulfide</td>
 +
            </tr>
 +
          <tr>
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          <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>
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            <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>
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            </tr>
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            <tr>
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            <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>
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          </table>
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        </div>
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      </div>
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          <div id="h2">
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        <div align="center">
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          <img src="https://static.igem.org/mediawiki/2017/0/09/T-Nanjing-China-project-h2.png" width="30%"/>
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          <table width="80%" border="0" cellspacing="1" cellpadding="1">
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          <tr>
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            <td colspan="2">
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              <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" />
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              </td>
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            </tr>
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            <tr>
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            <td colspan="2">
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              <p>The sequence was a good detecter in the lab work.</p>
 +
              </td></tr>
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            <tr>
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              <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>
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              <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>
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              <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>
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        </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>
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Revision as of 04:21, 28 October 2017

Team:Nanjing-China - 2017.igem.org

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.

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.

In order to demonstrate the design, a lot of experiments have been done.

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)

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)
It is worth to be mentioned that the team OUC help us demonstrate the result.

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.
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.
b) A visible photograph of a).
c) Test of selectivity.

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

Fluorescence intensity remains stationary when IPTG is added.
And Fluorescence intensity increases in a low hydrogen atmosphere.
When the amount of hydrogen goes to a higher level Fluorescence intensity increases apparently. meaning the designed reporter pathway have worked.

Figure 3. Influence of H2 concentration on fluorescence expression

Address

Life Science Department
#163 Xianlin Blvd, Qixia District
Nanjing University
Nanjing, Jiangsu Province
P.R. of China
Zip: 210046

Email

nanjing_china@163.com

Social media

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iGEM Nanjing-China@nanjing_china