Team:Tokyo Tech/Parts

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<title>Tokyo_Tech</title>
<title>Tokyo_Tech</title>
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<img src="https://static.igem.org/mediawiki/2014/thumb/b/b7/Tokyo_Tech_Logo.png/613px-Tokyo_Tech_Logo.png" height="150" /></h1></div>
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        <div align="right"><img src="https://static.igem.org/mediawiki/2014/b/b4/Tokyo_Tech_Logo2.png" height="150" /></div>
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<a href="https://2014.igem.org/Main_Page"><img src="https://static.igem.org/mediawiki/2014/thumb/8/84/Tokyo_Tech_iGEM_logo.png/794px-Tokyo_Tech_iGEM_logo.png" width="90" /></a></div>
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        <a href="http://www.titech.ac.jp/english/index.html" id="logo-link"></a>
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<ul>
<ul>
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<li><a href="#">Experiment</a>
<li><a href="#">Experiment</a>
  <ul>
  <ul>
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        <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Experiment/Prhl_reporter_assay" style="width:400px; margin-left:-135px;">Prhl reporter assay </a></li>
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        <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Experiment/Plux and Prhl reporter assay" style="width:400px; margin-left:-135px;">Plux and Prhl reporter assay</a></li>
 +
        <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Experiment/Prhl_reporter_assay" style="width:400px; margin-left:-135px;">Improved Prhl reporter assay</a></li>
        <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Experiment/C4HSL-dependent_3OC12HSL_production" style="width:400px; margin-left:-135px;">C4HSL-dependent 3OC12HSL production</a></li>
        <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Experiment/C4HSL-dependent_3OC12HSL_production" style="width:400px; margin-left:-135px;">C4HSL-dependent 3OC12HSL production</a></li>
        <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Experiment/3OC12HSL-dependent_C4HSL_production" style="width:400px; margin-left:-135px;">3OC12HSL-dependent C4HSL production</a></li>
        <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Experiment/3OC12HSL-dependent_C4HSL_production" style="width:400px; margin-left:-135px;">3OC12HSL-dependent C4HSL production</a></li>
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        <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Experiment/Symbiosis_confirmation_by_co-culture" style="width:400px; margin-left:-135px;">Symbiosis confirmation by co-culture </a></li>
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        <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Experiment/Symbiosis_confirmation_by_co-culture" style="width:400px; margin-left:-135px;">Mutualism Confirmation ~Co-culture Assay~</a></li>
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        <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Experiment/Transporter_assay" style="width:400px; margin-left:-135px;">Transporter assay</a></li>
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  </ul>
  </ul>
</li>
</li>
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<li><a href="https://2014.igem.org/Team:Tokyo_Tech/Modeling">Modeling</a></li>
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<li><a href="#">Modeling</a>
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                          <ul>
 +
                              <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Modeling/Overview"  style="width:400px; margin-left:-135px;">Overview</a></li>
 +
                              <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Modeling/Growth Conditions For Company And Customer"  style="width:400px; margin-left:-135px;">Growth Conditions For Company And Customer</a></li>
 +
                              <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Modeling/Analysis of C4HSL-dependent Switch" style="width:400px; margin-left:-135px;">Analysis of C4HSL-dependent Switch</a></li>
 +
                              <li><a href="https://2014.igem.org/Team:Tokyo_Tech/Modeling/Economic Wave"  style="width:400px; margin-left:-135px;">Economic Wave</a></li>
 +
                          </ul>
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                        </li>
<li class="current_page_item"><a href="https://2014.igem.org/Team:Tokyo_Tech/Parts">Parts</a></li>
<li class="current_page_item"><a href="https://2014.igem.org/Team:Tokyo_Tech/Parts">Parts</a></li>
<li><a href="https://2014.igem.org/Team:Tokyo_Tech/Policy_and_Practices" style="height:50px; padding-top:3px;">Policy and Practices</a></li>
<li><a href="https://2014.igem.org/Team:Tokyo_Tech/Policy_and_Practices" style="height:50px; padding-top:3px;">Policy and Practices</a></li>
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             <div class="post">
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      <h2 class="title"><a href="https://2014.igem.org/Team:Tokyo_Tech/Policy_and_Practice">Parts</a></h2>
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      <h2 class="title">Parts</h2>
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             <span class="meta">Each part give us a whole new experience </span>
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             <span class="meta">Each part gives us a whole new experience </span>
             <div class="entry-long">
             <div class="entry-long">
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              <div id="gototop"><a href="#"><img src="https://static.igem.org/mediawiki/2014/5/55/Tokyo_Tech_Go-to-top-icon.png" height="50" /></a></div>
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               <h3>&nbsp;</h3>
               <h3>&nbsp;</h3>
               <h2>Favorite Tokyo Tech 2014 iGEM Team Parts
               <h2>Favorite Tokyo Tech 2014 iGEM Team Parts
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                 <tr class="underName">
                 <tr class="underName">
                   <td>&nbsp;</td>
                   <td>&nbsp;</td>
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                   <td><a href="http://parts.igem.org/Part:BBa_K1529301">K1529301</a></td>
+
                   <td><a href="http://parts.igem.org/Part:BBa_K1529301">BBa_K1529301</a></td>
                   <td>Composite</td>
                   <td>Composite</td>
                   <td>Prhl(RL)-GFP</td>
                   <td>Prhl(RL)-GFP</td>
                   <td>Kohdai Hibi</td>
                   <td>Kohdai Hibi</td>
-
                   <td>801 </td>
+
                   <td>990 </td>
                 </tr>
                 </tr>
                 <tr class="underName">
                 <tr class="underName">
                   <td>&nbsp;</td>
                   <td>&nbsp;</td>
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                   <td><a href="http://parts.igem.org/Part:BBa_K1529265">K1529265</a></td>
+
                   <td><a href="http://parts.igem.org/Part:BBa_K1529302">BBa_K1529302</a></td>
                   <td>Composite</td>
                   <td>Composite</td>
-
                   <td>Plux-CmR-RhlI</td>
+
                   <td>Prhl(RL)-CmR-LasI</td>
                   <td>Kohdai Hibi</td>
                   <td>Kohdai Hibi</td>
                   <td>1435</td>
                   <td>1435</td>
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                 <tr class="underName">
                 <tr class="underName">
                   <td>&nbsp;</td>
                   <td>&nbsp;</td>
-
                   <td><a href="http://parts.igem.org/Part:BBa_K1529263">K1529263</a></td>
+
                   <td><a href="http://parts.igem.org/Part:BBa_K1529797">BBa_K1529797</a></td>
                   <td>Composite</td>
                   <td>Composite</td>
-
                   <td>Plux-CmR-LasI</td>
+
                   <td>Plux-CmR-RhlI</td>
-
                   <td>Kohdai Hibi</td>
+
                   <td>Shoko Suzuki </td>
                   <td>1435</td>
                   <td>1435</td>
                 </tr>
                 </tr>
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                 <p>&nbsp;</p>
                 <p>&nbsp;</p>
               </div>
               </div>
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               <h2>Tokyo Tech 2013 iGEM Team Parts</h2>
+
               <h2>Tokyo Tech 2014 iGEM Team Parts</h2>
               <div align="center">
               <div align="center">
                 <table width="799" border="1">
                 <table width="799" border="1">
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                   <tr class="underName">
                   <tr class="underName">
                     <td>&nbsp;</td>
                     <td>&nbsp;</td>
-
                     <td><a href="http://parts.igem.org/Part:BBa_K1529311">K1529311</a></td>
+
                     <td><a href="http://parts.igem.org/Part:BBa_K1529311">BBa_K1529311</a></td>
                     <td>Composite</td>
                     <td>Composite</td>
-
                     <td>Prhl(RL)-GFP</td>
+
                     <td>Prhl(LR)-GFP</td>
                     <td>Kohdai Hibi</td>
                     <td>Kohdai Hibi</td>
-
                     <td>801 </td>
+
                     <td>990 </td>
                   </tr>
                   </tr>
                   <tr class="underName">
                   <tr class="underName">
                     <td>&nbsp;</td>
                     <td>&nbsp;</td>
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                     <td><a href="http://parts.igem.org/Part:BBa_K1529321">K1529321</a></td>
+
                     <td><a href="http://parts.igem.org/Part:BBa_K1529321">BBa_K1529321</a></td>
                     <td>Composite</td>
                     <td>Composite</td>
-
                     <td>Prhl(a)-GFP</td>
+
                     <td>Prhl(RR)-GFP</td>
                     <td>Kohdai Hibi</td>
                     <td>Kohdai Hibi</td>
-
                     <td>801</td>
+
                     <td>990</td>
-
                  </tr>
+
-
                  <tr class="underName">
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-
                    <td>&nbsp;</td>
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-
                    <td><a href="http://parts.igem.org/Part:BBa_K1529264">K1529264</a></td>
+
-
                    <td>Composite</td>
+
-
                    <td>Plux-CmR-LuxI</td>
+
-
                    <td>Kohdai Hibi</td>
+
-
                    <td>1411</td>
+
-
                  </tr>
+
-
                  <tr class="underName">
+
-
                    <td>&nbsp;</td>
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-
                    <td><a href="http://parts.igem.org/Part:BBa_K1529302">K1529302</a></td>
+
-
                    <td>Composite</td>
+
-
                    <td>Prhl(RL)_CmR_LasI</td>
+
-
                    <td>Kohdai Hibi</td>
+
-
                    <td>1435</td>
+
-
                  </tr>
+
-
                  <tr class="underName">
+
-
                    <td>&nbsp;</td>
+
-
                    <td><a href="http://parts.igem.org/Part:BBa_K1529020">K1529020</a></td>
+
-
                    <td>Composite</td>
+
-
                    <td>Ptet-ydeD-tolC</td>
+
-
                    <td>Kohdai Hibi</td>
+
-
                    <td>1633</td>
+
                   </tr>
                   </tr>
 +
                 
                 </table>
                 </table>
                 <p>&nbsp;</p>
                 <p>&nbsp;</p>
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                 <tr>
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                   <td colspan="2"><h2>Best new Basic part<span class="head">: <a href="http://parts.igem.org/Part:BBa_K1529300">BBa_K1529300</a>, <a href="http://parts.igem.org/Part:BBa_K1529310">BBa_K1529310</a>, <a href="http://parts.igem.org/Part:BBa_K1529320">BBa_K1529320</a></span></h2></td>
+
                   <td><h2>1. Best New Basic Part<span class="head">:<a href="http://parts.igem.org/Part:BBa_K1529301"> BBa_K1529301</a>, <a href="http://parts.igem.org/Part:BBa_K1529311">BBa_K1529311</a>, <a href="http://parts.igem.org/Part:BBa_K1529321">BBa_K1529321</a></span></h2></td>
                 </tr>
                 </tr>
                 <tr>
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                   <td width="561"><p class="head">&#8226; Prhl(RL)  : <a href="http://parts.igem.org/Part:BBa_K1529300">BBa_K1529300</a> <br />
+
                   <td><p class="head">&#8226; Prhl(RL)-GFP (<a href="http://parts.igem.org/Part:BBa_K1529301">BBa_K1529301</a><a href="http://parts.igem.org/Part:BBa_K1529300"></a>)</p></td>
-
                    &#8226; Prhl(LR)  : <a href="http://parts.igem.org/Part:BBa_K1529310">BBa_K1529310</a> <br />
+
-
                  &#8226; Prhl(RR)  : <a href="http://parts.igem.org/Part:BBa_K1529320">BBa_K1529320</a> </p>
+
-
                    <p align="justify" class="info-18">Prhl(RL) Promoter has the biggest ratio  of leakage and expression (when C4HSL is induced) among the three above.</p>                  </td>
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                  <td width="329"><img src="http://cyberd.org/img/3/Under-Construction-Anime.jpg" height="300" /></td>
+
                 </tr>
                 </tr>
                 <tr>
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                   <td>&nbsp;</td>
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                   <td><p class="info-18">Prhl(RL) is a promoter that is activated by C4HSL-RhlR complex.&nbsp;<br />
-
                  <td>&nbsp;</td>
+
                    We improved Prhl promoter (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_R0071">BBa_R0071</a>) by changing LuxR binding site of Plux promoter (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_R0062">BBa_R0062</a>) to RhlR binding site. <br />
 +
                    To characterize the function of the&nbsp;Prhl(RL) promoter (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K1529300">BBa_K1529300</a>), we constructed this part,  Prhl(RL)-GFP (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K1529301">BBa_K1529301</a>) , by inserting a Prhl(RL) promoter into the upstream region of GFP coding sequence . </p>
 +
                    <p class="info-18">By using reporter cells containing Prhl(RL)-GFP, we measured the fluorescence intensity of the cells induced by C4HSL.&nbsp;<br />
 +
                    We confirmed that our new&nbsp;Prhl(RL)&nbsp;promoter was actually activated by C4HSL through an induction assay. (Fig. 5-1-1-1.)
 +
                  </p></td>
                 </tr>
                 </tr>
</table>
</table>
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  <table width="900" border="0">
                 <tr>
                 <tr>
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                   <td colspan="2"><h2>Best new Composite part<span class="head">: <a href="http://parts.igem.org/Part:BBa_K1529301">BBa_K1529301</a>, <a href="http://parts.igem.org/Part:BBa_K1529302">BBa_K1529302</a>, <a href="http://parts.igem.org/Part:BBa_K1529797">BBa_K1529797</a><a href="http://parts.igem.org/Part:BBa_K1139020" rel="nofollow"></a></span></h2></td>
+
                   <td><div align="center"><a href="https://2014.igem.org/File:Tokyo_Tech_Fig5-1-1-1.png"><img src="https://static.igem.org/mediawiki/2014/2/20/Tokyo_Tech_Fig5-1-1-1.png" width="709" height="370" /></a></div></td>
                 </tr>
                 </tr>
                 <tr>
                 <tr>
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                   <td colspan="2"><p class="head">Prhl(RL)_GFP  (<a href="http://parts.igem.org/Part:BBa_K1529301">BBa_K1529301</a>)</p></td>
+
                   <td><div align="center"><strong>Fig. 5-1-1-1.</strong> Fluorescence intensity detected by flow cytometer </div></td>
                 </tr>
                 </tr>
                 <tr>
                 <tr>
-
                   <td colspan="2"><p align="justify" class="info-18">Prhl(RL)  is a promoter that is activated by C4HSL.
+
                   <td>&nbsp;</td>
-
                    We improved Prhl Promoter(<a href="http://parts.igem.org/wiki/index.php/Part:BBa_R0071">BBa_R0071</a>)  by changing LuxR binding site of Plux Promoter(<a href="http://parts.igem.org/wiki/index.php/Part:BBa_R0062">BBa_R0062</a>) to  RhlR binding site.
+
-
                    To characterize the function of the&nbsp;Prhl(RL)  Promoter (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K1529300">BBa_K1529300</a>), we  constructed this part,  Prhl(RL)_GFP (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K1529301">BBa_K1529301</a>) , by  inserting a Prhl(RL) promoter into the upstream region of GFP coding sequence. By  using reporter cells containing Prhl(RL)_GFP, we measured the fluorescence  intensity of the cells induced by C4HSL.&nbsp;We confirmed that our new&nbsp;Prhl(RL)&nbsp;promoter  was actually activated by C4HSL through an induction assay (Fig. 5-1-1).</p>                  </td>
+
                 </tr>
                 </tr>
                 <tr>
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                   <td colspan="2"><div align="center"></div>                     <div align="center"><img src="" height="400" /></div></td>
+
                   <td><h2>2. Best New Composite Part<span class="head">: <a href="http://parts.igem.org/Part:BBa_K1529302">BBa_K1529302</a>, <a href="http://parts.igem.org/Part:BBa_K1529797">BBa_K1529797</a></span></h2></td>
                 </tr>
                 </tr>
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                <tr>
+
              <td><p class="head">&#8226;Prhl(RL)-CmR-LasI (<a href="http://parts.igem.org/Part:BBa_K1529302">BBa_K1529302</a>)</p></td>
-
                  <td colspan="2"><div align="center">                     Fig. 5-1-1.   Prhl(RL) has no leakage and higher expression</div>                    <div align="center"></div></td>
+
                 </tr>
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 +
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 +
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                   <td width="445">&nbsp;</td>
+
                   <td><p class="info-18">We constructed this part by combining &nbsp;<a href="http://parts.igem.org/Part:BBa_K1529300">BBa_K1529300</a>,&nbsp;<a href="http://parts.igem.org/Part:BBa_K395160">BBa_K395160</a>,&nbsp;<a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>&nbsp;and&nbsp;<a href="http://parts.igem.org/Part:BBa_C0078">BBa_C0078</a>.&nbsp;Prhl(RL) promoter has no leakage and higher expression than Prhl promoter (<a href="http://parts.igem.org/Part:BBa_R0071">BBa_R0071</a>). CmR and LasI are inserted into the downstream of the promoter</p>
-
                  <td width="445">&nbsp;</td>
+
                    <p class="info-18"This is the first BioBrick part that succeeded in C4HSL dependent CmR and 3OC12HSL production. Using this part with the plasmid that is constitutively expressing RhlR, we succeeded in confirming RhlR activates the expression of CmR and LasI in the presence of C4HSL. </p>
 +
                    <p class="info-18">
 +
To confirm CmR production, we added chloramphenicol into the medium containing Prhl(RL)-CmR-LasI cell and measured optical density for about 8 h to estimate the concentration of the cell. (Fig. 5-1-2-1.)
 +
</p>
 +
                  <p class="info-18"> Moreover, by co-culturing the cells containing this part with the cells containing 3OC12HSL dependent C4HSL producer part (<a href="http://parts.igem.org/Part:BBa_K1529797">BBa_K1529797</a>), we succeeded in constructing a positive feedback system.                    </p></td>
                 </tr>
                 </tr>
                 <tr>
                 <tr>
-
                   <td colspan="2"><p class="head">Prhl(RL)_CmR_LasI  (<a href="http://parts.igem.org/Part:BBa_K1529302">BBa_K1529302</a>)</p></td>
+
                   <td><div align="center"><a href="https://2014.igem.org/File:Tokyo_Tech_Fig5-1-2-1.png"><img src="https://static.igem.org/mediawiki/2014/1/1d/Tokyo_Tech_Fig5-1-2-1.png" height="400" /></a></div></td>
                 </tr>
                 </tr>
                 <tr>
                 <tr>
-
                   <td colspan="2"><p align="justify" class="info-18">We  constructed this part by combining&nbsp;<a href="http://parts.igem.org/Part:BBa_K1529300">BBa_K1529300</a>, <a href="http://parts.igem.org/Part:BBa_K395160">BBa_K395160</a>, <a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>&nbsp;and&nbsp;<a href="http://parts.igem.org/Part:BBa_C0078">BBa_C0078</a>. <br />
+
                   <td><div align="center"> <strong>Fig. 5-1-2-1. </strong>C4HSL-Dependent CmR Expression Assay</div>                     <div align="center"></div></td>
-
                    (CmR is Chloramphenicol  resistance.)
+
-
                  </p>
+
-
                    <p align="justify" class="info-18">Prhl(RL)  Promoter has no leakage and higher expression than Prhl Promoter (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_R0071">BBa_R0071</a>). CmR  and LasI are inserted into the downstream of the promoter.
+
-
                      This is the first Biobrick part  that succeeded in C4HSL dependent CmR and 3OC12HSL production.  Using this part with the plasmid that is constitutively expressing RhlR, we  succeeded in confirming RhlR activates the expression of CmR and LasI in the  presence of C4HSL.</p>
+
-
                    <p align="justify" class="info-18">Moreover, by co-culturing the  cells containing this part with the cells containing 3OC12HSL dependent C4HSL  producer part (<a href="http://parts.igem.org/Part:BBa_K1529797">BBa_K1529797</a>), we succeeded in constructing a positive  feedback system.</p>                    </td>
+
-
                </tr>
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-
                <tr>
+
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                  <td colspan="2"><div align="center"></div>
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                      <div align="center"><img src="" height="400" /></div></td>
+
-
                </tr>
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                <tr>
+
-
                  <td colspan="2"><div align="center"> Fig. 5-1-2.   </div>
+
-
                      <div align="center"></div></td>
+
                 </tr>
                 </tr>
                 <tr>
                 <tr>
-
                  <td>&nbsp;</td>
 
                   <td>&nbsp;</td>
                   <td>&nbsp;</td>
                 </tr>
                 </tr>
                 <tr>
                 <tr>
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                   <td colspan="2"><p class="head">Plux_CmR_RhlI (<a href="http://parts.igem.org/Part:BBa_K1529797">BBa_K1529797</a>)</p></td>
+
                 
 +
 
 +
<tr>
 +
                   <td><p class="head">&#8226; Plux-CmR-RhlI (<a href="http://parts.igem.org/Part:BBa_K1529797">BBa_K1529797</a>)</p></td>
                 </tr>
                 </tr>
                 <tr>
                 <tr>
-
                   <td colspan="2"><p class="info-18">We constructed this part by combining&nbsp;<a href="http://parts.igem.org/Part:BBa_K395162">BBa_K395162</a>,&nbsp;<a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>&nbsp;and&nbsp;<a href="http://parts.igem.org/Part:BBa_C0070">BBa_C0070</a>. <br />
+
                   <td><p class="info-18">We constructed this part by combining&nbsp;<a href="http://parts.igem.org/Part:BBa_K395162">BBa_K395162</a>,&nbsp;<a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>&nbsp;and&nbsp;<a href="http://parts.igem.org/Part:BBa_C0070">BBa_C0070</a>. (CmR is Chloramphenicol resistance.) <br />
-
                    (CmR is Chloramphenicol resistance.)  
+
                  This is the first BioBrick part that succeeded in 3OC12HSL dependent CmR and C4HSL production. In our experiment, we used 3OC12HSL instead of 3OC6HSL as an inducer in order to circumvent crosstalk. Using this part with the plasmid that is constitutively expressing LuxR, we succeeded in confirming LuxR activates the expression of CmR and RhlI in the presence of 3OC12HSL. </p>
-
                    </p>
+
                  <p class="info-18">To confirm C4HSL production, we measured the expression of GFP in reporter cells by flow cytometer. (Fig. 5-1-2-2).</p></td>
-
                    <p class="info-18">This is the first Biobrick part that succeeded in 3OC12HSL dependent CmR and C4HSL production. In our experiment, we used 3OC12HSL instead of 3OC6HSL as an inducer in order to circumvent crosstalk. Using this part with the plasmid that is constitutively expressing luxR, we succeeded in confirming LuxR activates the expression of CmR and RhlI in the presence of 3OC12HSL.</p></td>
+
                 </tr>
                 </tr>
                 <tr>
                 <tr>
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                   <td colspan="2"><div align="center"></div>
+
                   <td><div align="center"></div>
-
                       <div align="center"><img src="" height="400" /></div></td>
+
                       <div align="center"><a href="https://2014.igem.org/File:Tokyo_Tech_Fig5-1-2-2.png"><img src="https://static.igem.org/mediawiki/2014/e/e6/Tokyo_Tech_Fig5-1-2-2.png" height="400" /></a></div></td>
                 </tr>
                 </tr>
                 <tr>
                 <tr>
-
                   <td colspan="2"><div align="center"> Fig. 5-1-3.  </div>
+
                   <td><div align="center"><strong>Fig. 5-1-2-2.</strong> 3OC12HSL-Dependent C4HSL Production Assay</div>
                       <div align="center"></div></td>
                       <div align="center"></div></td>
                 </tr>
                 </tr>
                 <tr>
                 <tr>
                   <td>&nbsp;</td>
                   <td>&nbsp;</td>
-
                   <td>&nbsp;</td>
+
                </tr>
 +
                   <td><h2>3. Best  Part Collection: <span class="head"><a href="http://parts.igem.org/Part:BBa_K1529302">BBa_K1529302</a>, <a href="http://parts.igem.org/Part:BBa_K1529797">BBa_K1529797</a><a href="http://parts.igem.org/Part:BBa_K1139201" rel="nofollow"></a></span> </h2></td>
                 </tr>
                 </tr>
                 <tr>
                 <tr>
-
                   <td colspan="2"><h2>Best  Part Collection:<span class="head"> <a href="http://parts.igem.org/Part:BBa_K1529302">BBa_K1529302</a>, <a href="http://parts.igem.org/Part:BBa_K1529797">BBa_K1529797</a>, <a href="http://parts.igem.org/Part:BBa_K1529263">BBa_K1529263</a><a href="http://parts.igem.org/Part:BBa_K1139201" rel="nofollow"></a></span> </h2></td>
+
                   <td><p class="info-18">Using these parts enable us to accomplish mutualism. The results of the co-culture assay is shown in Fig. 5-1-2-3. Company’s characteristics are C4HSL-dependent survival and 3OC12HSL production (<a href="http://parts.igem.org/Part:BBa_K1529302">BBa_K1529302</a>), and Customer’s characteristics are the opposite from Company’s (<a href="http://parts.igem.org/Part:BBa_K1529797">BBa_K1529797</a>). From these characteristics, the symbiosis between the two cells can be established.</p></td>
 +
                </tr>
 +
                  <tr>
 +
                  <td><p class="info-18">Two types of fluorescent proteins were used to trace the growth of each cells in our symbiosis experiments. We constructed the Company cell containing GFP and Customer cell containing RFP. By measuring the OD of the cells expressing GFP with flow cytometer, the symbiosis was detected.</p><td>
 +
                  </tr>
 +
                  <tr>
 +
                  <td><p class="info-18">Using these parts enable us to accomplish mutualism. The results of the co-culture assay is shown in Fig. 5-1-2-3. Company’s By looking at the fluorescence intensity of GFP in Company cells, the optical density increased faster  in co-culture than single culture.From this point, we can say that Company and Customer actually mutualize in the medium.
 +
</p></td>
                 </tr>
                 </tr>
                 <tr>
                 <tr>
-
                   <td colspan="2"><p class="head">Plux_CmR_LasI (<a href="http://parts.igem.org/Part:BBa_K1529263">BBa_K1529263</a>)</p></td>
+
                   <td><div align="center"><a href="https://2014.igem.org/File:Tokyo_Tech_Fig5-1-2-3.png"><img src="https://static.igem.org/mediawiki/2014/0/08/Tokyo_Tech_Fig5-1-2-3.png" height="400" /></a></div></td>
                 </tr>
                 </tr>
                 <tr>
                 <tr>
-
                   <td colspan="2"><p class="info-18">We  constructed this part by combining&nbsp;<a href="http://parts.igem.org/Part:BBa_K395162">BBa_K395162</a>,&nbsp;<a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>&nbsp;and&nbsp;<a href="http://parts.igem.org/Part:BBa_C0078">BBa_C0078</a>.<br />
+
                   <td><div align="center"><strong>Fig. 5-1-2-3.</strong> The result of co-culture assay </div></td>
-
                    (CmR is Chloramphenicol  resistance.)</p>
+
-
                    <p class="info-18">To characterize Plux-CmR-LasI  (<a href="http://parts.igem.org/Part:BBa_K1529263">BBa_K1529263</a>), we introduced Plux-CmR-LasI on pSB3K3 with Ptet-RhlR-Ptet-GFP  on pSB6A1 to E.coli as “C4HSL dependent CmR and 3OC12HSL producer cell”. In  this E.coli, constitutively expressed RhlR activates the expression of CmR and  LasI in the presence of C4HSL.                    </p>
+
-
                    <p class="info-18">&nbsp;</p></td>
+
                 </tr>
                 </tr>
                 <tr>
                 <tr>
-
                   <td colspan="2"><div align="center"></div>
+
                   <td>&nbsp;</td>
-
                      <div align="center"><img src="" height="400" /></div></td>
+
                 </tr>
                 </tr>
                 <tr>
                 <tr>
-
                   <td colspan="2"><div align="center"> Fig. 5-1-4.  </div>
+
                   <td>&nbsp;</td>
-
                      <div align="center"></div></td>
+
                 </tr>
                 </tr>
                 <tr>
                 <tr>
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                  <td>&nbsp;</td>
 
                   <td>&nbsp;</td>
                   <td>&nbsp;</td>
                 </tr>
                 </tr>
               </table>
               </table>
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              <h2>Our submitted parts</h2>
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<!--              <h2>Our submitted parts</h2>
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<td class="status_cell  "></td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_white">&nbsp;</td><td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1529301" class="noul_link part_link">BBa_K1529301</a></td><td>Composite</td><td>Prhl(RL)_GFP</td><td width="100px">Kohdai Hibi</td><td align="right">990</td>
+
<td class="status_cell  "></td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_white">&nbsp;</td><td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1529301" class="noul_link part_link">BBa_K1529301</a></td><td>Composite</td><td>Prhl(RL)-GFP</td><td width="100px">Kohdai Hibi</td><td align="right">990</td>
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<td class="status_cell  "></td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_white">&nbsp;</td><td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1529311" class="noul_link part_link">BBa_K1529311</a></td><td>Composite</td><td>Prhl(LR)_GFP</td><td width="100px">Kohdai Hibi</td><td align="right">990</td>
+
<td class="status_cell  "></td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_white">&nbsp;</td><td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1529311" class="noul_link part_link">BBa_K1529311</a></td><td>Composite</td><td>Prhl(LR)-GFP</td><td width="100px">Kohdai Hibi</td><td align="right">990</td>
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<td class="status_cell  "></td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_white">&nbsp;&nbsp;</td><td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1529321" class="noul_link part_link">BBa_K1529321</a></td><td>Composite</td><td>Prhl(RR)_GFP</td><td width="100px">Kohdai Hibi</td><td align="right">990</td>
+
<td class="status_cell  "></td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_white">&nbsp;&nbsp;</td><td><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1529321" class="noul_link part_link">BBa_K1529321</a></td><td>Composite</td><td>Prhl(RR)-GFP</td><td width="100px">Kohdai Hibi</td><td align="right">990</td>
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<script>$('#groupparts').load('/cgi/api/groupparts.cgi', { t:'iGEM014', g:'Tokyo_Tech' },function(){ $('#groupparts .tablesorter').tablesorter();} );</script> -->
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<groupparts>iGEM014 Tokyo_Tech</groupparts>
 

Latest revision as of 03:58, 18 October 2014

Tokyo_Tech

Parts

Each part gives us a whole new experience

 

Favorite Tokyo Tech 2014 iGEM Team Parts

 
Name
Type
Description
Design
Length (bp)
  BBa_K1529301 Composite Prhl(RL)-GFP Kohdai Hibi 990
  BBa_K1529302 Composite Prhl(RL)-CmR-LasI Kohdai Hibi 1435
  BBa_K1529797 Composite Plux-CmR-RhlI Shoko Suzuki 1435

 

Tokyo Tech 2014 iGEM Team Parts

 
Name
Type
Description
Design
Length (bp)
  BBa_K1529311 Composite Prhl(LR)-GFP Kohdai Hibi 990
  BBa_K1529321 Composite Prhl(RR)-GFP Kohdai Hibi 990

 

1. Best New Basic Part: BBa_K1529301, BBa_K1529311, BBa_K1529321

• Prhl(RL)-GFP (BBa_K1529301)

Prhl(RL) is a promoter that is activated by C4HSL-RhlR complex. 
We improved Prhl promoter (BBa_R0071) by changing LuxR binding site of Plux promoter (BBa_R0062) to RhlR binding site.
To characterize the function of the Prhl(RL) promoter (BBa_K1529300), we constructed this part,  Prhl(RL)-GFP (BBa_K1529301) , by inserting a Prhl(RL) promoter into the upstream region of GFP coding sequence .

By using reporter cells containing Prhl(RL)-GFP, we measured the fluorescence intensity of the cells induced by C4HSL. 
We confirmed that our new Prhl(RL) promoter was actually activated by C4HSL through an induction assay. (Fig. 5-1-1-1.)

Fig. 5-1-1-1. Fluorescence intensity detected by flow cytometer
 

2. Best New Composite Part: BBa_K1529302, BBa_K1529797

•Prhl(RL)-CmR-LasI (BBa_K1529302)

We constructed this part by combining  BBa_K1529300BBa_K395160BBa_B0034 and BBa_C0078. Prhl(RL) promoter has no leakage and higher expression than Prhl promoter (BBa_R0071). CmR and LasI are inserted into the downstream of the promoter

To confirm CmR production, we added chloramphenicol into the medium containing Prhl(RL)-CmR-LasI cell and measured optical density for about 8 h to estimate the concentration of the cell. (Fig. 5-1-2-1.)

Moreover, by co-culturing the cells containing this part with the cells containing 3OC12HSL dependent C4HSL producer part (BBa_K1529797), we succeeded in constructing a positive feedback system.

Fig. 5-1-2-1. C4HSL-Dependent CmR Expression Assay
 

• Plux-CmR-RhlI (BBa_K1529797)

We constructed this part by combining BBa_K395162BBa_B0034 and BBa_C0070. (CmR is Chloramphenicol resistance.)
This is the first BioBrick part that succeeded in 3OC12HSL dependent CmR and C4HSL production. In our experiment, we used 3OC12HSL instead of 3OC6HSL as an inducer in order to circumvent crosstalk. Using this part with the plasmid that is constitutively expressing LuxR, we succeeded in confirming LuxR activates the expression of CmR and RhlI in the presence of 3OC12HSL.

To confirm C4HSL production, we measured the expression of GFP in reporter cells by flow cytometer. (Fig. 5-1-2-2).

Fig. 5-1-2-2. 3OC12HSL-Dependent C4HSL Production Assay
 

3. Best Part Collection: BBa_K1529302, BBa_K1529797

Using these parts enable us to accomplish mutualism. The results of the co-culture assay is shown in Fig. 5-1-2-3. Company’s characteristics are C4HSL-dependent survival and 3OC12HSL production (BBa_K1529302), and Customer’s characteristics are the opposite from Company’s (BBa_K1529797). From these characteristics, the symbiosis between the two cells can be established.

Two types of fluorescent proteins were used to trace the growth of each cells in our symbiosis experiments. We constructed the Company cell containing GFP and Customer cell containing RFP. By measuring the OD of the cells expressing GFP with flow cytometer, the symbiosis was detected.

Using these parts enable us to accomplish mutualism. The results of the co-culture assay is shown in Fig. 5-1-2-3. Company’s By looking at the fluorescence intensity of GFP in Company cells, the optical density increased faster in co-culture than single culture.From this point, we can say that Company and Customer actually mutualize in the medium.

Fig. 5-1-2-3. The result of co-culture assay