Team:LZU-China

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  <p><img src="lzuchianmfctt.jpg" width="1366" height="541" /></p>
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  <p>&nbsp;</p>
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  <p><img src="https://static.igem.org/mediawiki/2014/b/bc/Lzumfctt2.jpg" width="1366" height="542" /></p>
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  <p><img src="https://static.igem.org/mediawiki/2014/c/c6/Lzuchianmfctt.jpg" width="1366" height="541" /></p>
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  <p>&nbsp;</p>
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  <div>CONTENTS</div>
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  <p>&nbsp;</p>
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  <p>&nbsp;</p>
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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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      <p>&nbsp;</p></td>
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      <td width="12%"><p><a href="https://2014.igem.org/Team:LZU-China/background"><img src="https://static.igem.org/mediawiki/2014/8/83/1backgroud.png" width="100" height="100" /></a></p>
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      <p>Background</p></td>
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      <td width="12%"><p><a href="https://2014.igem.org/Team:LZU-China/wetlab"><img src="https://static.igem.org/mediawiki/2014/d/d5/2wetlab.png" width="100" height="100" /></a></p>
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      <p>Wet Lab</p></td>
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      <td width="12%"><p><a href="https://2014.igem.org/Team:LZU-China/drylab"><img src="https://static.igem.org/mediawiki/2014/c/c6/3drylab.png" width="100" height="100" /></a></p>
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      <p>Dry Lab</p></td>
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       <td width="12%"><p><a href="https://2014.igem.org/Team:LZU-China/Parts"><img src="https://static.igem.org/mediawiki/2014/1/14/4parts.png" width="100" height="100" /></a></p>
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      <p>Parts</p></td>
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      <td width="12%"><p><a href="https://2014.igem.org/Team:LZU-China/hp"><img src="https://static.igem.org/mediawiki/2014/5/5f/5humanpractics.png" width="100" height="100" /></a></p>
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      <p>Human Practice</p></td>
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      <td width="12%"><p><a href="https://2014.igem.org/Team:LZU-China/Team"><img src="https://static.igem.org/mediawiki/2014/a/a2/6members.png" width="100" height="100" /></a></p>
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      <p>Our team</p></td>
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      <td width="12%"><p><a href="https://2014.igem.org/Team:LZU-China/interlab"><img src="https://static.igem.org/mediawiki/2014/c/cc/7interlab.png" width="100" height="100" /></a></p>
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      <p>Interlab</p></td>
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      <td width="12%"><p><a href="https://2014.igem.org/Team:LZU-China/Safety"><img src="https://static.igem.org/mediawiki/2014/e/ee/8safety.png" width="100" height="100" /></a></p>
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      <p>Safety</p></td>
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      <td width="12%"><p><a href="https://2014.igem.org/Team:LZU-China/Notebook"><img src="https://static.igem.org/mediawiki/2014/0/00/9notebook.png" width="100" height="100" /></a></p>
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      <p>Notebook</p></td>
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      <td width="12%"><p><a href="https://2014.igem.org/Team:LZU-China/future"><img src="https://static.igem.org/mediawiki/2014/0/04/10future.png" width="100" height="100" /></a></p>
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      <p>Future Work</p></td>
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      <td width="20%">&nbsp;</td>
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       <td><div> *In order to get better user experience, we recommend the Internet Explorer 10 (or higher version). And the <span id="tran_1_0" data-aligning="#src_1_0,#tran_1_0" jquery171013312911573948127="19">best</span> <span id="tran_1_1" data-aligning="#src_1_1,#tran_1_1" jquery171013312911573948127="16">resolution</span> is 1366 X 768.</div>
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      <td width="49%"><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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       <div> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;GENETIC ENGINEERED BACTERIA</div>
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       <div> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;ABSTRACT</div>
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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;
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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;Microbial Fuel Cells (MFCs) which can convert contaminants in wastewaters into energy is an ideal approach to solve both pollution problem and energy crisis. However, MFCs still have disadvantages such as hard to determine the contaminants concentrations which is a major drawback for MFCs applications. <br />
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            <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>
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             &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;In our study, we found that the electricity produced by MFCs was somewhat related to the substrates concentrations such as P-nitrophenol (PNP) in anode or Chromium (VI) in cathode. Therefore, we hypothesize that a) by using genetic engineered bacteria, the MFC's electricity will be more stable and correlated with substrates concentrations. b) by monitoring MFC's electricity, it will be possible to measure substrates concentrations. <br />
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            <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>
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             &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;In this study, we've designed a novel MFC system. 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. In the cathode, gene codes chromate (VI) reductase Yief was cloned into E.coli. The stability of MFCs has been improved and the electricity generated to correlated with the substrates. Moreover, based on this correlation, we've designed a program which is able to monitor the contaminants concentrations in MFCs. <br /></td>
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            <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>
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            <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Besides, we find many  genes can catalyze the reduction of Cr(VI). They can be used in the cathode to reduce these heavy metal ions.</p>
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             <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;<img src="https://static.igem.org/mediawiki/2014/3/39/WETAimage002.png" width="556" height="307" /></p>
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            <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>
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      <td ><div>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;ABOUT THE CONSTRUCTION OF PNP SENSOR</div></td>
 
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        <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).
 
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          We got a good result.You can see the PNP  can also induce the green fluorescence.</p>
 
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      <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;<img src="https://static.igem.org/mediawiki/2014/3/3c/Image005.png" width="455" height="340" /></p>
 
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      <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;Figure-3 Fluorescence  of different system. </p>
 
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      <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;a.bacterial liquid with 10mM PNP;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;b.bacterial liquid with  ddH2O;</p>
 
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      <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;c.bacterial liquid with 10mM KNO3;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;d.bacterial  liquid with 10mM KCl.</p>
 
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      <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Firstly we got the plasmid with K1172303  from Registry. We cut the plasmid by EcoRI and XbaI, then put the <em>Pnsr</em>(K216005+B0030  this sequence was synthesized by company)  into the gap.</p>
 
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      <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;<img src="https://static.igem.org/mediawiki/2014/0/06/Image008.jpg" width="489" height="90" /></p>
 
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      <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;Figure-4  Construction of K1523101</p>
 
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      <p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The size of the whole part is about  3700bp(without plasmid), we test the assembling result by PCR(sense:5&rsquo;---TTCCCATCTATAATCCTCCCTGATTCTTCG---3&rsquo;;anti-sense:5&rsquo;---GAATTCTCTAGATTACAACTGTTGTTCAAGCTGTT---3&rsquo;).  From the gel picture we can see the size is right.</p>
 
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      <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;<img src="https://static.igem.org/mediawiki/2014/c/c1/Image009.png" width="643" height="305" /></p>
 
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      <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;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Figure-5  Gel picture of K1523101&rsquo;s PCR</p>
 
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      <td background="https://static.igem.org/mediawiki/2014/a/a6/Huils.jpg" ><div>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;ABOUT THE REDUCING Cr IONS GENES</div></td>
 
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        <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>
 
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        <p align="left"><strong><em>nahG</em></strong>: We got the sequence from <em>Pseudomonas putida</em> </p>
 
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  <strong><em>nahE: </em></strong>We got the sequence from <em>Pseudomonas putida<strong> </strong></em><br />
 
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          &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(Primer:sense:5&rsquo;---CCGGAATTCGCGGCCGCTTCTAGATGTCGAATAAAATTATGAAAACGTCGCG---3&rsquo;;<br />
 
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  <strong><em>nahR: </em></strong>We got the sequence from <em>Pseudomonas putida<strong> </strong></em><br />
 
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          &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(Primer:sense:5&rsquo;---  CCGGAATTCGCGGCCGCTTCTAGATGAAAAACAATAAACCTGGCTTGCGC---3&rsquo;;<br />
 
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  <strong><em>chrR: </em></strong>We got the sequence from <em>Escherichia coli str.  K-12<strong> </strong></em><br />
 
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  <strong><em>yieF: </em></strong>We got the sequence from <em>Escherichia coli str.  K-12<strong> </strong></em><br />
 
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          &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;(Primer:sense:5&rsquo;---  GAATTCGGATCCATGTCTGAAAAATTGCAGGTGG---3&rsquo;;<br />
 
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Revision as of 14:30, 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

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

CONTENTS

 

 

 

 

 

Background

Wet Lab

Dry Lab

Parts

Human Practice

 
 

 

 

 

   

 

   
 

Our team

Interlab

Safety

Notebook

Future Work

 

 

 

 

 

 

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        ABSTRACT
              Microbial Fuel Cells (MFCs) which can convert contaminants in wastewaters into energy is an ideal approach to solve both pollution problem and energy crisis. However, MFCs still have disadvantages such as hard to determine the contaminants concentrations which is a major drawback for MFCs applications.
           In our study, we found that the electricity produced by MFCs was somewhat related to the substrates concentrations such as P-nitrophenol (PNP) in anode or Chromium (VI) in cathode. Therefore, we hypothesize that a) by using genetic engineered bacteria, the MFC's electricity will be more stable and correlated with substrates concentrations. b) by monitoring MFC's electricity, it will be possible to measure substrates concentrations.
           In this study, we've designed a novel MFC system. 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. In the cathode, gene codes chromate (VI) reductase Yief was cloned into E.coli. The stability of MFCs has been improved and the electricity generated to correlated with the substrates. Moreover, based on this correlation, we've designed a program which is able to monitor the contaminants concentrations in MFCs.