Difference between revisions of "Team:INSA-Lyon/Composite Part"

 
(15 intermediate revisions by 3 users not shown)
Line 70: Line 70:
 
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Proof">Proof of Concept</a></li>
 
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Proof">Proof of Concept</a></li>
 
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Composite_Part">Parts</a></li>
 
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Composite_Part">Parts</a></li>
 +
                <li><a href="https://2016.igem.org/Team:INSA-Lyon/Notebook">Notebook</a></li>
 
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Achievements">Achievements</a></li>
 
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Achievements">Achievements</a></li>
 
               </ul>
 
               </ul>
Line 77: Line 78:
 
               <ul class="dropdown-menu" role="menu">
 
               <ul class="dropdown-menu" role="menu">
 
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Experiments">Protocols</a></li>
 
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Experiments">Protocols</a></li>
−
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Demonstrate">Results</a></li>
+
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Results">Results</a></li>
 
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Safety">Safety</a></li>
 
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Safety">Safety</a></li>
−
                <li><a href="https://2016.igem.org/Team:INSA-Lyon/Notebook">Notebook</a></li>
 
 
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Interlab">Interlab</a></li>
 
                 <li><a href="https://2016.igem.org/Team:INSA-Lyon/Interlab">Interlab</a></li>
 
               </ul>
 
               </ul>
Line 122: Line 122:
 
   <div class="col-md-3 contents-sidebar">
 
   <div class="col-md-3 contents-sidebar">
 
       <ul id="sidebar" class="nav nav-stacked hidden-xs hidden-sm" data-spy="affix" data-target=".contents-sidebar" data-offset="100">
 
       <ul id="sidebar" class="nav nav-stacked hidden-xs hidden-sm" data-spy="affix" data-target=".contents-sidebar" data-offset="100">
−
          <li><a href="#characptac">Characterization of pTAC</a></li>
 
 
           <li><a href="#transcriptase">HIV reverse transcriptase</a></li>
 
           <li><a href="#transcriptase">HIV reverse transcriptase</a></li>
 
           <li><a href="#streptavidin">Streptavidin chimeric protein</a></li>
 
           <li><a href="#streptavidin">Streptavidin chimeric protein</a></li>
 +
          <li><a href="#characptac">Characterization of pTAC</a></li>
 
       </ul>
 
       </ul>
 
   </div>
 
   </div>
Line 136: Line 136:
 
                       <div class="row">
 
                       <div class="row">
 
                             <div class="col-md-2 col-md-offset-5"><h5 class="wow fadeInUp" data-wow-duration="0.5s" data-wow-delay="0.5s">PARTS</h5></div>
 
                             <div class="col-md-2 col-md-offset-5"><h5 class="wow fadeInUp" data-wow-duration="0.5s" data-wow-delay="0.5s">PARTS</h5></div>
−
                             <div class="col-md-2 col-md-offset-3"><a class="read-more-btn wow fadeInUp" data-wow-duration="1s" data-wow-delay="1s" href="#summary">Read More</a></div>
+
                             <div class="col-md-2 col-md-offset-3"><a class="read-more-btn wow fadeInUp" data-wow-duration="1s" data-wow-delay="1s" href="#parts">Read More</a></div>
 
                           </div>
 
                           </div>
 
                         <span class="line"></span>
 
                         <span class="line"></span>
Line 166: Line 166:
 
                                   The aim of our project is to create a device to detect Sexually Transmitted Infections (STIs). To that end, we have created three series of BioBricks that enabled us to investigate and characterize the function of different pieces of our self-test.
 
                                   The aim of our project is to create a device to detect Sexually Transmitted Infections (STIs). To that end, we have created three series of BioBricks that enabled us to investigate and characterize the function of different pieces of our self-test.
 
                               </p>
 
                               </p>
−
                               <div class="subtitle" id="characptac">
+
                                
−
                                  <h2 class="title">Characterization of pTAC promoter</h2>
+
−
                                  <span class="line"></span>
+
−
                                  <p>
+
−
                                      Using synthetic biology principles, we have created a self-diagnostic device to detect several Sexually Transmitted Infections (STIs), starting with  two major health threats, the Human Immunodeficiency Virus (HIV) and the Hepatitis B Virus (HBV), as an alternative to sampling and laboratory testing.
+
−
                                  </p>
+
−
                                  <h2>
+
−
                                      <a href="http://parts.igem.org/Part:BBa_K1934040">BBa_K1934040</a>: RFP under control of pTAC promoter
+
−
                                  </h2>
+
−
                                  <p>
+
−
                                    This composite part contains the coding sequence of the RFP reporter gene downstream of pTAC and strong RBS sequences.
+
−
                                  </p>
+
−
 
+
−
                                  <img src="https://static.igem.org/mediawiki/2016/e/ee/T--INSA-Lyon--parts1.png" class="img-responsive center-block">
+
−
 
+
−
 
+
−
                                  <h2>
+
−
                                      <a href="http://parts.igem.org/Part:BBa_K1934050">BBa_K1934050</a>: CFP under control of pTAC promoter.
+
−
                                  </h2>
+
−
                                  <p>
+
−
                                      This composite part contains the coding sequence of the CFP reporter gene downstream of pTAC and strong RBS sequences.
+
−
                                  </p>
+
−
 
+
−
                                  <img src="https://static.igem.org/mediawiki/2016/f/fd/T--INSA-Lyon--parts2.png" class="img-responsive center-block">
+
−
                              </div>
+
  
  
Line 199: Line 175:
 
                                       The objective of our project is to detect some Sexually Transmitted Infections (STIs) and especially, HIV with the HIV reverse transcriptase and HBV with its surface antigen HBsAg. In order to test our detection device, we created two composite parts which contain the coding sequences of p51 and p66 subunits of HIV reverse transcriptase respectively.
 
                                       The objective of our project is to detect some Sexually Transmitted Infections (STIs) and especially, HIV with the HIV reverse transcriptase and HBV with its surface antigen HBsAg. In order to test our detection device, we created two composite parts which contain the coding sequences of p51 and p66 subunits of HIV reverse transcriptase respectively.
 
                                   </p>
 
                                   </p>
−
                                   <h2>
+
                                   <h4>
 
                                   <a href="http://parts.igem.org/Part:BBa_K1934060">BBa_K1934060</a>: p51 subunit  of HIV reverse transcriptase.
 
                                   <a href="http://parts.igem.org/Part:BBa_K1934060">BBa_K1934060</a>: p51 subunit  of HIV reverse transcriptase.
−
                                   </h2>
+
                                   </h4>
 
                                   <p>
 
                                   <p>
−
                                       This composite part contains the coding sequence of the p51 subunit under the control of T7 promoter, a RBS and a T7 terminator.
+
                                       This composite part contains the coding sequence of the p51 subunit (<a href="http://www.uniprot.org/uniprot/P03366">UniProtKB P03366</a>, <a href="http://www.rcsb.org/pdb/explore/explore.do?structureId=5d3g">PDB 5D3G_B</a>), which directly comes from the genome of the <a href="https://www.ncbi.nlm.nih.gov/nuccore/9629357?report=genbank">HIV 1 group M subtype B</a>, under the control of T7 promoter, a RBS and a T7 terminator.
  
 
                                   <img src="https://static.igem.org/mediawiki/2016/b/b3/T--INSA-Lyon--parts3.png" class="img-responsive center-block">
 
                                   <img src="https://static.igem.org/mediawiki/2016/b/b3/T--INSA-Lyon--parts3.png" class="img-responsive center-block">
 
                                   </p>
 
                                   </p>
  
−
                                   <h2>
+
                                   <h4>
 
                                   <a href="http://parts.igem.org/Part:BBa_K1934061">BBa_K1934061</a>: p66 subunit of HIV reverse transcriptase.
 
                                   <a href="http://parts.igem.org/Part:BBa_K1934061">BBa_K1934061</a>: p66 subunit of HIV reverse transcriptase.
−
                                   </h2>
+
                                   </h4>
−
                                   <p>This composite part contains the coding sequence of the p66 subunit under the control of T7 promoter, a RBS and a T7 terminator.
+
                                   <p>This composite part contains the coding sequence of the p66 subunit (<a href="http://www.uniprot.org/uniprot/P03366">UniProtKB P03366</a>, <a href="http://www.rcsb.org/pdb/explore/explore.do?structureId=5d3g">PDB 5D3G_A</a>), which directly comes from the genome of the <a href="https://www.ncbi.nlm.nih.gov/nuccore/9629357?report=genbank">HIV 1 group M subtype B</a>,  under the control of T7 promoter, a RBS and a T7 terminator.
 
                                   </p>
 
                                   </p>
 
                                   <img src="https://static.igem.org/mediawiki/2016/f/f0/T--INSA-Lyon--parts4.png" class="img-responsive center-block">
 
                                   <img src="https://static.igem.org/mediawiki/2016/f/f0/T--INSA-Lyon--parts4.png" class="img-responsive center-block">
Line 222: Line 198:
 
                                       As an anchor, we chose to use the streptavidin protein combined with one or two cellulose binding domain(s) in order to be easily fixed onto a cellulose paper.
 
                                       As an anchor, we chose to use the streptavidin protein combined with one or two cellulose binding domain(s) in order to be easily fixed onto a cellulose paper.
 
                                   </p>
 
                                   </p>
−
                                   <h2>
+
                                   <h4>
 
                                       <a href="http://parts.igem.org/Part:BBa_K1934020">BBa_K1934020</a>: Streptavidin with Cellulose Binding Domains (CBDs).
 
                                       <a href="http://parts.igem.org/Part:BBa_K1934020">BBa_K1934020</a>: Streptavidin with Cellulose Binding Domains (CBDs).
−
                                   </h2>
+
                                   </h4>
 
                                   <p>This composite part contains the coding sequence of the streptavidin protein combined with two CBDs, which corresponds to the Stanford-Brown 2014 iGEM team part <a href="http://parts.igem.org/Part:BBa_K1499004">(BBa_K1499004)</a> , under the control of a pTAC promoter, a strong RBS and a bidirectional terminator.
 
                                   <p>This composite part contains the coding sequence of the streptavidin protein combined with two CBDs, which corresponds to the Stanford-Brown 2014 iGEM team part <a href="http://parts.igem.org/Part:BBa_K1499004">(BBa_K1499004)</a> , under the control of a pTAC promoter, a strong RBS and a bidirectional terminator.
 
                                   </p>
 
                                   </p>
 
                                   <img src="https://static.igem.org/mediawiki/2016/7/7d/T--INSA-Lyon--parts5.png" class="img-responsive center-block">
 
                                   <img src="https://static.igem.org/mediawiki/2016/7/7d/T--INSA-Lyon--parts5.png" class="img-responsive center-block">
  
−
                                   <h2>
+
                                   <h4>
 
                                       <a href="http://parts.igem.org/Part:BBa_K1934000">BBa_K1934000</a>: RFP with Cellulose Binding Domains (CBDs)
 
                                       <a href="http://parts.igem.org/Part:BBa_K1934000">BBa_K1934000</a>: RFP with Cellulose Binding Domains (CBDs)
−
                                   </h2>
+
                                   </h4>
 
                                   <p>To verify the efficacy of the CBDs for binding to the cellulose, we created this composite part in which the coding sequences for two different CBDs were fused respectively upstream and downstream of the coding sequence for RFP.
 
                                   <p>To verify the efficacy of the CBDs for binding to the cellulose, we created this composite part in which the coding sequences for two different CBDs were fused respectively upstream and downstream of the coding sequence for RFP.
 
                                   </p>
 
                                   </p>
Line 237: Line 213:
 
                                   <img src="https://static.igem.org/mediawiki/2016/c/cf/T--INSA-Lyon--parts6.png" class="img-responsive">
 
                                   <img src="https://static.igem.org/mediawiki/2016/c/cf/T--INSA-Lyon--parts6.png" class="img-responsive">
  
−
                                   <h2>
+
                                   <h4>
 
                                       <a href="http://parts.igem.org/Part:BBa_K1934030 center-block">BBa_K1934030</a>: Streptavidin with CBD-CipA (Cellulosomal scaffolding protein A)
 
                                       <a href="http://parts.igem.org/Part:BBa_K1934030 center-block">BBa_K1934030</a>: Streptavidin with CBD-CipA (Cellulosomal scaffolding protein A)
−
                                   </h2>
+
                                   </h4>
−
                                   <p>In order to test another type of CBDs, we created this composite part which presents the coding sequence of streptavidin followed by the coding sequence of CBD-CipA (Cellulosomal scaffolding protein A) protein. We have submitted this part to the <strong>“Best new composite part” special prize.</strong></p>
+
                                   <p>In order to test another type of CBDs, we created this composite part which presents the coding sequence of streptavidin followed by the coding sequence of CBD-CipA (Cellulosomal scaffolding protein A) protein. As CBD-CipA shows a better interaction with cellulose, this part is an <strong>improvement of the streptavidin with classical CBDs  <a href="http://parts.igem.org/Part:BBa_K1499004">(BBa_K1499004)</a></strong>. We have submitted this part to the <strong>“Best new composite part” special prize.</strong></p>
  
 
                                   <img src="https://static.igem.org/mediawiki/2016/6/6c/T--INSA-Lyon--parts7.png" class="img-responsive center-block">
 
                                   <img src="https://static.igem.org/mediawiki/2016/6/6c/T--INSA-Lyon--parts7.png" class="img-responsive center-block">
 +
                              </div>
 +
 +
                              <div class="subtitle" id="characptac">
 +
                                  <h2 class="title">Characterization of pTAC promoter</h2>
 +
                                  <span class="line"></span>
 +
                                  <p>
 +
                                    pTAC is a promoter that we used in our different composite parts (eg for surproduction of proteins). As it’s induced by IPTG, by linking a reporter gene downstream of pTAC, the two following parts allowed us to characterize the promoter's responsiveness to IPTG, to quantify the promoter leakage and thus to <strong>improve the basic part containing the pTAC promoter sequence <a href="http://parts.igem.org/Part:BBa_K864400">(BBa_K864400)</a></strong>.
 +
                                  </p>
 +
                                  <h4>
 +
                                      <a href="http://parts.igem.org/Part:BBa_K1934040">BBa_K1934040</a>: RFP under control of pTAC promoter
 +
                                  </h4>
 +
                                  <p>
 +
                                    This composite part contains the coding sequence of the RFP reporter gene downstream of pTAC and strong RBS sequences.
 +
                                  </p>
 +
 +
                                  <img src="https://static.igem.org/mediawiki/2016/e/ee/T--INSA-Lyon--parts1.png" class="img-responsive center-block">
 +
 +
 +
                                  <h4>
 +
                                      <a href="http://parts.igem.org/Part:BBa_K1934050">BBa_K1934050</a>: CFP under control of pTAC promoter.
 +
                                  </h4>
 +
                                  <p>
 +
                                      This composite part contains the coding sequence of the CFP reporter gene downstream of pTAC and strong RBS sequences.
 +
                                  </p>
 +
 +
                                  <img src="https://static.igem.org/mediawiki/2016/f/fd/T--INSA-Lyon--parts2.png" class="img-responsive center-block">
 
                               </div>
 
                               </div>
  
Line 259: Line 261:
 
    
 
    
 
    
 
    
−
    <!-- Start footer -->
+
  <!-- Start footer -->
 
   <footer id="footer">
 
   <footer id="footer">
 
       <div class="container">
 
       <div class="container">
 
           <div class="row">
 
           <div class="row">
−
               <div class="col-md-6 col-sm-6">
+
            <div class="col-md-1 col-sm-1">
 +
              <img src="https://static.igem.org/mediawiki/2016/7/78/T--INSA-Lyon--Logo_IA2C.png">
 +
            </div>
 +
              <div class="col-md-1 col-md-offset-1 col-sm-1">
 +
              <a href="https://fondation.insa-lyon.fr/"><img src="https://static.igem.org/mediawiki/2016/c/cc/T--INSA-Lyon--Logo_Fondation_INSA.png"></a>
 +
              </div>
 +
              <div class="col-md-1 col-md-offset-1 col-sm-1">
 +
              <a href="http://www.biomerieux.com/"><img src="https://static.igem.org/mediawiki/2016/e/ea/T--INSA-Lyon--Logo_biomerieux.png"></a>
 +
              </div>
 +
              <div class="col-md-1 col-md-offset-1 col-sm-1">
 +
              <a href="https://www.france-science.org/"><img src="https://static.igem.org/mediawiki/2016/7/77/T--INSA-Lyon--Logo_Embassy_OST.jpg"></a>
 +
              </div>
 +
              <div class="col-md-1 col-md-offset-1 col-sm-1">
 +
              <a href="https://www.insa-lyon.fr/en/"><img src="https://static.igem.org/mediawiki/2016/9/99/T--INSA-Lyon--Logo_INSA_Lyon.jpg"></a>
 +
 
 +
          </div>
 +
          <p></br></p>
 +
          <p></br></p>
 +
 
 +
 
 +
          <div class="row">
 +
               <div class="col-md-8 col-sm-8">
 
                   <div class="footer-left">
 
                   <div class="footer-left">
 
                       <p>Designed by Team iGEM INSA-LYON </p>
 
                       <p>Designed by Team iGEM INSA-LYON </p>
 
                   </div>
 
                   </div>
 
               </div>
 
               </div>
−
               <div class="col-md-6 col-sm-6">
+
               <div class="col-md-4 col-sm-4">
 
                   <div class="footer-right">
 
                   <div class="footer-right">
 
                       <a href="https://www.facebook.com/insaadnconceptclub/"><i class="fa fa-facebook"></i></a>
 
                       <a href="https://www.facebook.com/insaadnconceptclub/"><i class="fa fa-facebook"></i></a>
 
                       <a href="http://www.dailymotion.com/iGEM_Lyon_2011"><i class="fa fa-video-camera"></i></a>
 
                       <a href="http://www.dailymotion.com/iGEM_Lyon_2011"><i class="fa fa-video-camera"></i></a>
−
                       <a href="https://www.linkedin.com/groups/8265587"><i class="fa fa-linkedin"></i></a>
+
                       <a href="https://www.linkedin.com/groups/8265587"><i class="fa fa-linkedin"></i></a>
−
                       <a href="ia2c.asso@gmail.com"><i class="fa fa-envelope"></i></a>              
+
                       <a href="mailto:ia2c.asso@gmail.com"><i class="fa fa-envelope"></i></a>
 
                   </div>
 
                   </div>
 
               </div>
 
               </div>
 
           </div>
 
           </div>
 +
 
       </div>
 
       </div>
 
   </footer>
 
   </footer>
 
   <!-- End footer -->
 
   <!-- End footer -->
 +
  
  

Latest revision as of 22:34, 8 November 2016

iGEM : Parts

 
PARTS
Read More

Summary

The aim of our project is to create a device to detect Sexually Transmitted Infections (STIs). To that end, we have created three series of BioBricks that enabled us to investigate and characterize the function of different pieces of our self-test.

One of the targets: HIV reverse transcriptase

The objective of our project is to detect some Sexually Transmitted Infections (STIs) and especially, HIV with the HIV reverse transcriptase and HBV with its surface antigen HBsAg. In order to test our detection device, we created two composite parts which contain the coding sequences of p51 and p66 subunits of HIV reverse transcriptase respectively.

BBa_K1934060: p51 subunit of HIV reverse transcriptase.

This composite part contains the coding sequence of the p51 subunit (UniProtKB P03366, PDB 5D3G_B), which directly comes from the genome of the HIV 1 group M subtype B, under the control of T7 promoter, a RBS and a T7 terminator.

BBa_K1934061: p66 subunit of HIV reverse transcriptase.

This composite part contains the coding sequence of the p66 subunit (UniProtKB P03366, PDB 5D3G_A), which directly comes from the genome of the HIV 1 group M subtype B, under the control of T7 promoter, a RBS and a T7 terminator.

The anchor: streptavidin chimeric protein

As an anchor, we chose to use the streptavidin protein combined with one or two cellulose binding domain(s) in order to be easily fixed onto a cellulose paper.

BBa_K1934020: Streptavidin with Cellulose Binding Domains (CBDs).

This composite part contains the coding sequence of the streptavidin protein combined with two CBDs, which corresponds to the Stanford-Brown 2014 iGEM team part (BBa_K1499004) , under the control of a pTAC promoter, a strong RBS and a bidirectional terminator.

BBa_K1934000: RFP with Cellulose Binding Domains (CBDs)

To verify the efficacy of the CBDs for binding to the cellulose, we created this composite part in which the coding sequences for two different CBDs were fused respectively upstream and downstream of the coding sequence for RFP.

BBa_K1934030: Streptavidin with CBD-CipA (Cellulosomal scaffolding protein A)

In order to test another type of CBDs, we created this composite part which presents the coding sequence of streptavidin followed by the coding sequence of CBD-CipA (Cellulosomal scaffolding protein A) protein. As CBD-CipA shows a better interaction with cellulose, this part is an improvement of the streptavidin with classical CBDs (BBa_K1499004). We have submitted this part to the “Best new composite part” special prize.

Characterization of pTAC promoter

pTAC is a promoter that we used in our different composite parts (eg for surproduction of proteins). As it’s induced by IPTG, by linking a reporter gene downstream of pTAC, the two following parts allowed us to characterize the promoter's responsiveness to IPTG, to quantify the promoter leakage and thus to improve the basic part containing the pTAC promoter sequence (BBa_K864400).

BBa_K1934040: RFP under control of pTAC promoter

This composite part contains the coding sequence of the RFP reporter gene downstream of pTAC and strong RBS sequences.

BBa_K1934050: CFP under control of pTAC promoter.

This composite part contains the coding sequence of the CFP reporter gene downstream of pTAC and strong RBS sequences.



Designed by Team iGEM INSA-LYON