Difference between revisions of "Team:HUST-China/Collaborations"

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             <div class="collapse navbar-collapse" id="navbar-collapse">
 
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                         <ul class="dropdown-menu">
 
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                             <li><a href="https://2017.igem.org/Team:HUST-China/project/background">Background</a></li>
 
                             <li><a href="https://2017.igem.org/Team:HUST-China/project/background">Background</a></li>
                             <li><a href="https://2017.igem.org/Team:HUST-China/Description">Description</a></li>                                        
+
                             <li><a href="https://2017.igem.org/Team:HUST-China/Description">Description</a></li>                                  
 
                             <li><a href="https://2017.igem.org/Team:HUST-China/Safety">Safety</a></li>
 
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                            <li><a href="https://2017.igem.org/Team:HUST-China/Demonstrate">Demonstrate</a></li>
 
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                             WETLAB <b class="caret"></b>
 
                             WETLAB <b class="caret"></b>
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                             MODELING
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                             MODEL
 
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                             PARTS
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                            <li><a href="https://2017.igem.org/Team:HUST-China/Basic part">Basic part</a></li>
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                            <li><a href="https://2017.igem.org/Team:HUST-China/Part collection">Part collection</a></li>
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                             HUMAN PRACTICES
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                             HP<b class="caret"></b>
 
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                            <li><a href="https://2017.igem.org/Team:HUST-China/Collaborations"> Collaborations</a></li>
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                        </ul>
 
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     </nav>
 
     </nav>
  
 
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     <div class="tab1" id="section0" style="width:100%;margin-top: 65px;">
     <div class="tab1" style="width:100%;margin-top: 65px;">
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             <h1 class="tab-h1" ><strong>「Attributions」</strong></h1>
             <h1 class="tab-h1" ><strong>「Experiment」</strong></h1>
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     </div>
 
     </div>
  
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                 <div class="collapse navbar-collapse" id="navbar-collapse">
 
                 <div class="collapse navbar-collapse" id="navbar-collapse">
 
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                     <li class="active"><a href="#section1">Sensing part</a></li>
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                     <li class="active"><a href="#section1">HUST-Cheering</a></li>
                     <li><a href="#section2">Recycling part</a></li>
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                     <li><a href="#section2">NCTU iGEM conference</a></li>
                     <li><a href="#section3">The whole circuit </a></li>
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                     <li><a href="#section3">Equipment Sharing </a></li>
 
                 </ul>
 
                 </ul>
 
                 </div>   
 
                 </div>   
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             <div class="col-sm-8">
 
             <div class="col-sm-8">
 
                 <div id="section1" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;">     
 
                 <div id="section1" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;">     
                     <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">Sensing part</h3>
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                     <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">HUST-Cheering! Conference the 2<sup>nd</sup>!</h3>
                    <p>
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                     <p style="padding:5px">
                        The sensing part is to achieve the awareness of the engineering bacteria on the rare earth ions in the environment.
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                      The three schools in Wuhan, HUST-China, WHU-China and HZAU-China, always keep in touch and having collaborations. This summer, we keep the tradition, we held conferences in July and share the idea of synthesis biology when NCTU Formosa came to join us. Moreover, we discussed about our project and get valuable suggestions from each other.
                    </p>
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                     </p>  
                    <h4><strong>①Amplify</strong> </h4>
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                    <img title="demo1" src="https://static.igem.org/mediawiki/2017/0/01/2017_HUST_China_HP_image1.png" alt="demo1" class="col-xs-8 col-xs-offset-2" style="padding: 10px 0px;">    
                     <p>
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                        pmrA-B-Fe、pmrC/GFP  gene through the pcr protocol
+
                    </p>
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                    <h4><strong>②Plasmid Construction</strong> </h4>
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                    <p>
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                      Coding sequences pmrA/pmrB and pmrC-GFP were cloned into the expression vector pBAD30 by restriction enzyme digestion and DNA ligation.
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                     </p>
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                    <div class="container experiment_circuit col-md-6 col-xs-6 col-md-offset-3 col-xs-offset-3">
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                        <img src="https://static.igem.org/mediawiki/2017/d/d5/2017_HUST_China_Experiment_image1.png" alt="experiment_circuit" class="img-responsive">
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                    </div>
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                    <div class="row col-xs-12">
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                        <p>
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                            Then we transformed these vectors into Ecoli BL21 cells.
+
                        </p>
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                        <h4><strong>③Detection</strong> </h4>
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                        <p>
+
                          Arabinose was used to induce expression of pmrABC system.
+
                        </p>
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                        <p>
+
                          We use the original pmrA-pmrB / Fe-pmrC-GFP circuit to test whether our
+
                          pmrABC system works in our plasmid, then the expression and efficacy
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                          of LBT1-12 were tested .
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                        </p>
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                    </div>
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                    <div class="table-responsive" style="padding: 10px 100px; text-align: center; font-size:14px;">
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                        <table class="table">
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                          <thead>
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                              <tr>
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                                <th style="text-align: center;">gene circuit</th>
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                                <th style="text-align: center;">vector</th>
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                                <th style="text-align: center;">description</th>
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                              </tr>
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                          </thead>
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                          <tbody>
+
                                  <tr>
+
                                    <td>pmrA-pmrB/Fe-pmrC-GFP</td>
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                                    <td>pBAD30</td>
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                                    <td>Test whether our pmrABC system works in our plasmid</td>
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                                  </tr>
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                                  <tr>
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                                    <td>pmrA-pmrB/LBT1-pmrC-GFP </td>
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                                    <td>pBAD30</td>
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                                    <td> Detecting effectiveness of protein LBT1</td>
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                                  </tr>
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                                  <tr>
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                                    <td>pmrA-pmrB/LBT2-pmrC-GFP</td>
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                                    <td>pBAD30 </td>
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                                    <td> Detecting effectiveness of protein LBT2</td>
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                                  </tr>
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                                  <tr>
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                                    <td>pmrA-pmrB/LBT3-pmrC-GFP</td>
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                                    <td>pBAD30 </td>
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                                    <td>Detecting effectiveness of protein LBT3</td>
+
                                  </tr>
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                                  <tr>
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                                    <td>pmrA-pmrB/LBT4-pmrC-GFP</td>
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                                    <td> pBAD30 </td>
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                                    <td>Detecting effectiveness of protein LBT4</td>
+
                                  </tr>
+
                                  <tr>
+
                                    <td>pmrA-pmrB/LBT5-pmrC-GFP</td>
+
                                    <td>pBAD30</td>
+
                                    <td>Detecting effectiveness of protein LBT5</td>
+
                                  </tr>
+
                                  <tr>
+
                                    <td>pmrA-pmrB/LBT6-pmrC-GFP </td>
+
                                    <td>pBAD30</td>
+
                                    <td> Detecting effectiveness of protein LBT6</td>
+
                                  </tr>
+
                                  <tr>
+
                                    <td>pmrA-pmrB/LBT7-pmrC-GFP</td>
+
                                    <td>pBAD30 </td>
+
                                    <td> Detecting effectiveness of protein LBT7</td>
+
                                  </tr>
+
                                  <tr>
+
                                    <td>pmrA-pmrB/LBT8-pmrC-GFP</td>
+
                                    <td>pBAD30 </td>
+
                                    <td>Detecting effectiveness of protein LBT8</td>
+
                                  </tr>
+
                                  <tr>
+
                                    <td>pmrA-pmrB/LBT9-pmrC-GFP</td>
+
                                    <td> pBAD30 </td>
+
                                    <td>Detecting effectiveness of protein LBT9</td>
+
                                  </tr>
+
                                  <tr>
+
                                    <td>pmrA-pmrB/LBT10-pmrC-GFP</td>
+
                                    <td>pBAD30 </td>
+
                                    <td> Detecting effectiveness of protein LBT10</td>
+
                                  </tr>
+
                                  <tr>
+
                                    <td>pmrA-pmrB/LBT11-pmrC-GFP</td>
+
                                    <td>pBAD30 </td>
+
                                    <td>Detecting effectiveness of protein LBT11</td>
+
                                  </tr>
+
                                  <tr>
+
                                    <td>pmrA-pmrB/LBT12-pmrC-GFP </td>
+
                                    <td> pBAD30 </td>
+
                                    <td>Detecting effectiveness of protein LBT12</td>
+
                                  </tr>
+
                               
+
                          </tbody>
+
                        </table>
+
                    </div>
+
 
                 </div>
 
                 </div>
 
  
 
                 <div id="section2" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;">  
 
                 <div id="section2" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;">  
                     <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Recycling part </h3>
+
                     <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">NCTU iGEM conference</h3>
                     <p>
+
                     <p style="padding:5px">
                            The recycling part is to capture the rare earth ions in the environment and to anchor our cells over the silicon net.
+
                        This year we also participate in NCTU iGEM conference. We shared ideas with different Asian countries, discussed the difficulties and fun as an iGEMer. And most importantly, we show our project and realized its shortages during the statement. The suggestion gave by Prof. Chen bring us a way to enhance our recycle system, which directly advance our project progress.
                    </p>
+
                    <h4><strong>①With appropriate primers, PCR was carried out to amplify target gene oprf-LBT1-12 and oprf-Si tag.</strong> </h4>
+
                    <div class="container experiment_circuit col-md-6 col-xs-6 col-md-offset-3 col-xs-offset-3">
+
                        <img src="https://static.igem.org/mediawiki/2017/7/73/2017_HUST_China_Experiment_image2.png" alt="experiment_circuit" class="img-responsive">
+
                    </div>
+
                    <h4 class="col-xs-12"><strong>②Then we constructed two vectors to achieve expression of LBT and Si-tag.</strong> </h4>
+
                    <div class="container experiment_circuit col-md-6 col-xs-6 col-md-offset-3 col-xs-offset-3">
+
                        <img src="https://static.igem.org/mediawiki/2017/7/78/2017_HUST_China_Experiment_image3.png" alt="experiment_circuit" class="img-responsive">
+
                    </div>
+
                    <p class="col-xs-12">
+
                        We cloned the sequence of oprf-LBT and Si-tag into the vector pET28α in order to make the sequence be tagged N-terminally with a 6×His tag. 6×His tag is an affinity tag which allows the tagged recombinant protein to be purified in the process of affinity purification.  
+
                    </p>
+
                    <h4><strong>③Detection</strong> </h4>
+
                    <p>
+
                      In the meanwhile,we added the FLAG tag to achieve that whether the LBTs and the Si tag was expressed on the cell membrane.  
+
 
                     </p>
 
                     </p>
 +
                    <img title="demo1" src="https://static.igem.org/mediawiki/2017/f/fc/2017_HUST_China_HP_img2.png" alt="demo1" class="col-xs-8 col-xs-offset-2" style="padding: 10px 0px;">             
 
                 </div>
 
                 </div>
  
 
                 <div id="section3" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;">         
 
                 <div id="section3" style="border: solid 1px #666; margin:5px;border-radius:10px;overflow: hidden;">         
                     <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Construction of the whole circuit</h3>
+
                     <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Equipment Sharing</h3>
                     <p>
+
                     <p style="padding:5px">
                       All of our part should be built in one plasmid. Therefore, we use endonuclease and ligase to  replace the GFP by Oprf-LBT and Oprf-Si tag.
+
                       As this year a 96-well plate is required for parts shipment. But we lost two of them by mistake, which cause great trouble to our plan. When heard about this, WHU-China gave one of their plate to us generously. Before part submitting fezzed, the lyophilizer in WHU-China and HZAU-China was not unfortunately broken. So, we share the experimental equipment to help them submitting all the samples.  
                     </p>
+
                     </p>  
                    <div class="container experiment_circuit col-md-6 col-xs-6 col-md-offset-3 col-xs-offset-3" style="padding: 5px;">
+
                        <img src="https://static.igem.org/mediawiki/2017/5/58/2017_HUST_China_Experiment_image4.png" alt="experiment_circuit" class="img-responsive">
+
                    </div>
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            <h3><strong>Acknowledgments:</strong></h3>
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            <img style="max-width: 100%; height:auto; display: block " src="https://static.igem.org/mediawiki/2017/8/88/2017_HUST_China_seven_logo.png" /> 
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Revision as of 07:00, 29 October 2017

Attributions

「Attributions」

HUST-Cheering! Conference the 2nd!

The three schools in Wuhan, HUST-China, WHU-China and HZAU-China, always keep in touch and having collaborations. This summer, we keep the tradition, we held conferences in July and share the idea of synthesis biology when NCTU Formosa came to join us. Moreover, we discussed about our project and get valuable suggestions from each other.

demo1

NCTU iGEM conference

This year we also participate in NCTU iGEM conference. We shared ideas with different Asian countries, discussed the difficulties and fun as an iGEMer. And most importantly, we show our project and realized its shortages during the statement. The suggestion gave by Prof. Chen bring us a way to enhance our recycle system, which directly advance our project progress.

demo1

Equipment Sharing

As this year a 96-well plate is required for parts shipment. But we lost two of them by mistake, which cause great trouble to our plan. When heard about this, WHU-China gave one of their plate to us generously. Before part submitting fezzed, the lyophilizer in WHU-China and HZAU-China was not unfortunately broken. So, we share the experimental equipment to help them submitting all the samples.

Acknowledgments: