Team:LZU-China/tempfortest

From 2014.igem.org

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      <td width="1201" height="265" rowspan="2"><p><img src="https://static.igem.org/mediawiki/2014/thumb/8/84/2.wetlab.top.jpg/800px-2.wetlab.top.jpg" width="1366" height="270" /></p></td>
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      <td width="100%" height="199" bgcolor="#187ADB">&nbsp;</td>
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  <p>&nbsp;</p>
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      <td width="30%">&nbsp;</td>
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      <td width="40%"><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;In wet lab, we designed a novel MFC device containing genetic engineered bacteria. For  anode strain, we&rsquo;ve cloned a PNP sensor sequence and a riboflavin into  Escherichia coli. The recombinants is able to detect PNP and produce riboflavin  to boost electrical generation when co-cultured with<em> Shewanella oneidensis. </em></p></td>
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      <td width="30%">&nbsp;</td>
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  <p>&nbsp;</p>
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  <p>&nbsp;</p>
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      <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg" height="550" valign="top"><p>&nbsp;</p>
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        <p>&nbsp;</p>
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            <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg"  height="50">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;GENETIC ENGINEERED BACTERIA%黑%</td>
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          </tr>
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          <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg" width="30%" height="1146"><p>&nbsp;</p></td>
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          <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg" width="57%">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;We designed this pathway for our purpose. As you can see, the NsrR is constitutive expressed in almost all E. coli orgnisms and it can repress the promoter PyeaR. In our project we found that this sensitive system is also adjusted to the PNP(p-nitrophenol). So we constructed this sensor pathway.</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The sensor will cause the expression of riboflavin, a kind of mediator which can induce the generation of current in MFCs, to show the appearance of PNP. We can estimate the concentration of PNP. </p>
 +
            <div><img src="https://static.igem.org/mediawiki/2014/3/3c/Image005.png" alt="" width="455" height="340" /></div>
 +
            <p>&nbsp;</p>
 +
            <p align="center">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</p>
 +
            <p align="center">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Figure-1  The pathway of PNP sensor</p>
 +
            <p>&nbsp;</p>
 +
<div><img src="https://static.igem.org/mediawiki/2014/3/3c/Image005.png" alt="" width="455" height="340" /></div>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Figure-1  The pathway of PNP sensor</p>
 +
                        <p>&nbsp;</p>
 +
<div><img src="https://static.igem.org/mediawiki/2014/3/3c/Image005.png" alt="" width="455" height="340" /></div>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Figure-1  The pathway of PNP sensor</p>
 +
                        <p>&nbsp;</p>
 +
<div><img src="https://static.igem.org/mediawiki/2014/3/3c/Image005.png" alt="" width="455" height="340" /></div>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Figure-1  The pathway of PNP sensor</p>
 +
                        <p>&nbsp;</p>
 +
<div><img src="https://static.igem.org/mediawiki/2014/3/3c/Image005.png" alt="" width="455" height="340" /></div>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Figure-1  The pathway of PNP sensor</p>
 +
           
 +
            <p>&nbsp;</p>
 +
<div><img src="https://static.igem.org/mediawiki/2014/3/3c/Image005.png" alt="" width="455" height="340" /></div>
 +
            <p>&nbsp;</p>
 +
            <p align="center">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Figure-2  The pathway of the Cr ion reductase(yieF as an example)</p></td>
 +
          <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg" width="13%">&nbsp;</td>
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  </table>
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</div>
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<div id="apDiv6"><img src="https://static.igem.org/mediawiki/2014/6/6e/Lzulogo.png" width="220" height="221" /></div>
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<div id="apDiv8">
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            <td>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;dxdsx%白%</td>
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  </table>
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      <td  width="30%">&nbsp;</td>
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      <td  ><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;        </p>
 +
        <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;We found that the part BBa_K381001 by  iGEM10_BCCS-Bristol is a sensor which can detect the appearance of nitrate and  nitrite. So we perfomed an experience by this part to see if this part can  detect PNP(p-Nitrophenol).
 +
          We got a good result.You can see the PNP  can also induce the green fluorescence.</p>
 +
      <p>&nbsp;</p>
 +
                  <p>&nbsp;</p>
 +
<div><img src="https://static.igem.org/mediawiki/2014/3/3c/Image005.png" alt="" width="455" height="340" /></div>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Figure-1  The pathway of PNP sensor</p>
 +
                        <p>&nbsp;</p>
 +
<div><img src="https://static.igem.org/mediawiki/2014/3/3c/Image005.png" alt="" width="455" height="340" /></div>
 +
            <p>&nbsp;</p>
 +
            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Figure-1  The pathway of PNP sensor</p>
 +
                        <p>&nbsp;</p>
 +
<div><img src="https://static.igem.org/mediawiki/2014/3/3c/Image005.png" alt="" width="455" height="340" /></div>
 +
            <p>&nbsp;</p>
 +
        <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Figure-1  The pathway of PNP sensor</p>
 +
      <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</p>
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<p>&nbsp;</p></td>
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      <td  width="13%">&nbsp;</td>
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      <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg"><p>&nbsp;</p>
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    <tr>
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      <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg" ><div>
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        <table width="100%" border="0">
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          <tr>
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            <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;dgfb%黑色%</td>
 +
          </tr>
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        <p>&nbsp;</p>
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      </div></td>
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  </table>
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  <table width="100%" border="0" >
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      <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg" width="30%">&nbsp;</td>
 +
      <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg" width="57%"><p align="left">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</p>
 +
        <p align="left">&nbsp;</p>
 +
        <p align="left">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;We found  four genes to reduce the Cr(VI) to Cr(III). They are<em> nahG, nahR, nahE, chrR</em> and <em>yieF. </em>We didn&rsquo;t do the deep research about them because of time  limitation. They can be used in the general plasmids such as pBR322. We just  made them to be the formation of Biobrick. More uses and functions need to be  demonstrated and found.</p>
 +
        <p align="left"><strong><em>nahG</em></strong>: We got the sequence from <em>Pseudomonas putida</em> </p>
 +
        <p align="left">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(Primer:sense:5&rsquo;---CCGGAATTCGCGGCCGCTTCTAGATGAAAAACAATAAACCTGGCTTGCGC---3&rsquo;;<br />
 +
          &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;anti-sense:5&rsquo;---AAAATACTAGTAGCGGCCGCTGCAGTCACCCTTGACGTAGCACACC---3&rsquo;)<br />
 +
  <strong><em>nahE: </em></strong>We got the sequence from <em>Pseudomonas putida<strong> </strong></em><br />
 +
          &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(Primer:sense:5&rsquo;---CCGGAATTCGCGGCCGCTTCTAGATGTCGAATAAAATTATGAAAACGTCGCG---3&rsquo;;<br />
 +
          &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;anti-sense:5&rsquo;---AAAATACTAGTAGCGGCCGCTGCAGCTACTTCAATTCATTACTGTATTTAGCGTG---3&rsquo;)<br />
 +
  <strong><em>nahR: </em></strong>We got the sequence from <em>Pseudomonas putida<strong> </strong></em><br />
 +
          &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(Primer:sense:5&rsquo;---  CCGGAATTCGCGGCCGCTTCTAGATGAAAAACAATAAACCTGGCTTGCGC---3&rsquo;;<br />
 +
          &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;anti-sense:5&rsquo;---AAAATACTAGTAGCGGCCGCTGCAGTCACCCTTGACGTAGCACACC---3&rsquo;)<br />
 +
  <strong><em>chrR: </em></strong>We got the sequence from <em>Escherichia coli str.  K-12<strong> </strong></em><br />
 +
          &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(Primer:sense:5&rsquo;---  CCGGAATTCGCGGCCGCTTCTAGATGAAAAAAATAGTCCAGTCGGAAGG---3&rsquo;;<br />
 +
          &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;anti-sense:5&rsquo;---AAAATACTAGTAGCGGCCGCTGCAGTCAGGCCTTTTTGTGCTGTTCAAC---3&rsquo;)<br />
 +
  <strong><em>yieF: </em></strong>We got the sequence from <em>Escherichia coli str.  K-12<strong> </strong></em><br />
 +
          &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(Primer:sense:5&rsquo;---  GAATTCGGATCCATGTCTGAAAAATTGCAGGTGG---3&rsquo;;<br />
 +
        &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;anti-sense:5&rsquo;---GAATTCTCTAGATTAGATCTTAACTCGCTGAATAAA---3&rsquo;)</p>
 +
        <div>&nbsp;<img src="https://static.igem.org/mediawiki/2014/3/3c/Image005.png" alt="" width="455" height="340" /></div>
 +
        <p align="left">&nbsp;</p>
 +
<p align="left">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(Primer:sense:5&rsquo;---  CCGGAATTCGCGGCCGCTTCTAGATGAAAAAAATAGTCCAGTCGGAAGG---3&rsquo;;<br />
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;anti-sense:5&rsquo;---AAAATACTAGTAGCGGCCGCTGCAGTCAGGCCTTTTTGTGCTGTTCAAC---3&rsquo;)<br />
 +
<strong><em>yieF: </em></strong>We got the sequence from <em>Escherichia coli str.  K-12<strong> </strong></em><br />
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(Primer:sense:5&rsquo;---  GAATTCGGATCCATGTCTGAAAAATTGCAGGTGG---3&rsquo;;<br />
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;anti-sense:5&rsquo;---GAATTCTCTAGATTAGATCTTAACTCGCTGAATAAA---3&rsquo;)</p>
 +
<div>&nbsp;<img src="https://static.igem.org/mediawiki/2014/3/3c/Image005.png" alt="" width="455" height="340" /></div>
 +
<p align="left">&nbsp;</p>
 +
<p align="left">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(Primer:sense:5&rsquo;---  CCGGAATTCGCGGCCGCTTCTAGATGAAAAAAATAGTCCAGTCGGAAGG---3&rsquo;;<br />
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;anti-sense:5&rsquo;---AAAATACTAGTAGCGGCCGCTGCAGTCAGGCCTTTTTGTGCTGTTCAAC---3&rsquo;)<br />
 +
<strong><em>yieF: </em></strong>We got the sequence from <em>Escherichia coli str.  K-12<strong> </strong></em><br />
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(Primer:sense:5&rsquo;---  GAATTCGGATCCATGTCTGAAAAATTGCAGGTGG---3&rsquo;;<br />
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;anti-sense:5&rsquo;---GAATTCTCTAGATTAGATCTTAACTCGCTGAATAAA---3&rsquo;)</p>
 +
<div>&nbsp;<img src="https://static.igem.org/mediawiki/2014/3/3c/Image005.png" alt="" width="455" height="340" /></div>
 +
<p align="left">&nbsp;</p>
 +
<p align="left"></p>
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<p align="left">&nbsp;</p>
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<p align="left">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<a href="https://2014.igem.org/Team:LZU-China/wetlab2"><img src="https://static.igem.org/mediawiki/2014/e/e2/Mfcdb.png" width="349" height="81" /></a></p>
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Latest revision as of 14:02, 17 October 2014

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" " http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd"> LZU-China 2014

 
 

 

 

          In wet lab, we designed a novel MFC device containing genetic engineered bacteria. For anode strain, we’ve cloned a PNP sensor sequence and a riboflavin into Escherichia coli. The recombinants is able to detect PNP and produce riboflavin to boost electrical generation when co-cultured with Shewanella oneidensis.

 

 

 

 

 

 

        GENETIC ENGINEERED BACTERIA%黑%

 

            

          We designed this pathway for our purpose. As you can see, the NsrR is constitutive expressed in almost all E. coli orgnisms and it can repress the promoter PyeaR. In our project we found that this sensitive system is also adjusted to the PNP(p-nitrophenol). So we constructed this sensor pathway.

          The sensor will cause the expression of riboflavin, a kind of mediator which can induce the generation of current in MFCs, to show the appearance of PNP. We can estimate the concentration of PNP.

 

          

                                                              Figure-1 The pathway of PNP sensor

 

 

                                                              Figure-1 The pathway of PNP sensor

 

 

                                                              Figure-1 The pathway of PNP sensor

 

 

                                                              Figure-1 The pathway of PNP sensor

 

 

                                                              Figure-1 The pathway of PNP sensor

 

 

                                                             Figure-2 The pathway of the Cr ion reductase(yieF as an example)

 

 

 

 

             dxdsx%白%
 

          

          We found that the part BBa_K381001 by iGEM10_BCCS-Bristol is a sensor which can detect the appearance of nitrate and nitrite. So we perfomed an experience by this part to see if this part can detect PNP(p-Nitrophenol). We got a good result.You can see the PNP can also induce the green fluorescence.

 

 

 

                                                              Figure-1 The pathway of PNP sensor

 

 

                                                              Figure-1 The pathway of PNP sensor

 

 

                                                              Figure-1 The pathway of PNP sensor

                              

 

 

 

 

 

 

         dgfb%黑色%

 

 

 

         

 

          We found four genes to reduce the Cr(VI) to Cr(III). They are nahG, nahR, nahE, chrR and yieF. We didn’t do the deep research about them because of time limitation. They can be used in the general plasmids such as pBR322. We just made them to be the formation of Biobrick. More uses and functions need to be demonstrated and found.

nahG: We got the sequence from Pseudomonas putida

          (Primer:sense:5’---CCGGAATTCGCGGCCGCTTCTAGATGAAAAACAATAAACCTGGCTTGCGC---3’;
          anti-sense:5’---AAAATACTAGTAGCGGCCGCTGCAGTCACCCTTGACGTAGCACACC---3’)
nahE: We got the sequence from Pseudomonas putida
          (Primer:sense:5’---CCGGAATTCGCGGCCGCTTCTAGATGTCGAATAAAATTATGAAAACGTCGCG---3’;
          anti-sense:5’---AAAATACTAGTAGCGGCCGCTGCAGCTACTTCAATTCATTACTGTATTTAGCGTG---3’)
nahR: We got the sequence from Pseudomonas putida
          (Primer:sense:5’--- CCGGAATTCGCGGCCGCTTCTAGATGAAAAACAATAAACCTGGCTTGCGC---3’;
          anti-sense:5’---AAAATACTAGTAGCGGCCGCTGCAGTCACCCTTGACGTAGCACACC---3’)
chrR: We got the sequence from Escherichia coli str. K-12
          (Primer:sense:5’--- CCGGAATTCGCGGCCGCTTCTAGATGAAAAAAATAGTCCAGTCGGAAGG---3’;
          anti-sense:5’---AAAATACTAGTAGCGGCCGCTGCAGTCAGGCCTTTTTGTGCTGTTCAAC---3’)
yieF: We got the sequence from Escherichia coli str. K-12
          (Primer:sense:5’--- GAATTCGGATCCATGTCTGAAAAATTGCAGGTGG---3’;
          anti-sense:5’---GAATTCTCTAGATTAGATCTTAACTCGCTGAATAAA---3’)

 

 

     (Primer:sense:5’--- CCGGAATTCGCGGCCGCTTCTAGATGAAAAAAATAGTCCAGTCGGAAGG---3’;
          anti-sense:5’---AAAATACTAGTAGCGGCCGCTGCAGTCAGGCCTTTTTGTGCTGTTCAAC---3’)
yieF: We got the sequence from Escherichia coli str. K-12
          (Primer:sense:5’--- GAATTCGGATCCATGTCTGAAAAATTGCAGGTGG---3’;
          anti-sense:5’---GAATTCTCTAGATTAGATCTTAACTCGCTGAATAAA---3’)

 

 

     (Primer:sense:5’--- CCGGAATTCGCGGCCGCTTCTAGATGAAAAAAATAGTCCAGTCGGAAGG---3’;
          anti-sense:5’---AAAATACTAGTAGCGGCCGCTGCAGTCAGGCCTTTTTGTGCTGTTCAAC---3’)
yieF: We got the sequence from Escherichia coli str. K-12
          (Primer:sense:5’--- GAATTCGGATCCATGTCTGAAAAATTGCAGGTGG---3’;
          anti-sense:5’---GAATTCTCTAGATTAGATCTTAACTCGCTGAATAAA---3’)