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

 
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A composite part is a functional unit of DNA consisting of two or more basic parts assembled together. <a href="http://parts.igem.org/wiki/index.php/Part:BBa_I13507">BBa_I13507</a> is an example of a composite part, consisting of an RBS, a protein coding region for a red fluorescent protein, and a terminator.
 
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<p>New composite BioBrick devices can be made by combining existing BioBrick Parts (like Inverters, Amplifiers, Smell Generators, Protein Balloon Generators, Senders, Receivers, Actuators, and so on).</p>
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<h4>Note</h4>
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<p>This page should list all the composite parts your team has made during your project. You must add all characterization information for your parts on the Registry. You should not put characterization information on this page.</p>
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          <li><a href="#transcriptase">HIV reverse transcriptase</a></li>
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          <li><a href="#streptavidin">Streptavidin chimeric protein</a></li>
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          <li><a href="#characptac">Characterization of pTAC</a></li>
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                          <div class="title-area" id=summary>
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                              <h2 class="title">Summary</h2>
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                              <span class="line"></span>
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                              <p>
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                                  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.
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                              </p>
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                              <div class="subtitle" id="transcriptase">
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                                  <h2 class="title">One of the targets: HIV reverse transcriptase</h2>
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                                  <span class="line"></span>
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                                  <p>
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                                      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.
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                                  </p>
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                                  <h4>
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                                  <a href="http://parts.igem.org/Part:BBa_K1934060">BBa_K1934060</a>: p51 subunit  of HIV reverse transcriptase.
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                                  </h4>
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                                  <p>
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                                      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.
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                                  <img src="https://static.igem.org/mediawiki/2016/b/b3/T--INSA-Lyon--parts3.png" class="img-responsive center-block">
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                                  </p>
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                                  <h4>
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                                  <a href="http://parts.igem.org/Part:BBa_K1934061">BBa_K1934061</a>: p66 subunit of HIV reverse transcriptase.
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                                  </h4>
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                                  <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.
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                                  </p>
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                                  <img src="https://static.igem.org/mediawiki/2016/f/f0/T--INSA-Lyon--parts4.png" class="img-responsive center-block">
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                              </div>
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                              <div class="subtitle" id="streptavidin">
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                                  <h2 class="title">The anchor: streptavidin chimeric protein</h2>
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                                  <span class="line"></span>
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                                  <p>
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                                      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.
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                                  </p>
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                                  <h4>
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                                      <a href="http://parts.igem.org/Part:BBa_K1934020">BBa_K1934020</a>: Streptavidin with Cellulose Binding Domains (CBDs).
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                                  </h4>
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                                  <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.
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                                  </p>
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                                  <img src="https://static.igem.org/mediawiki/2016/7/7d/T--INSA-Lyon--parts5.png" class="img-responsive center-block">
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                                  <h4>
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                                      <a href="http://parts.igem.org/Part:BBa_K1934000">BBa_K1934000</a>: RFP with Cellulose Binding Domains (CBDs)
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                                  </h4>
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                                  <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.
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                                  </p>
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                                  <img src="https://static.igem.org/mediawiki/2016/c/cf/T--INSA-Lyon--parts6.png" class="img-responsive">
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                                  <h4>
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                                      <a href="http://parts.igem.org/Part:BBa_K1934030 center-block">BBa_K1934030</a>: Streptavidin with CBD-CipA (Cellulosomal scaffolding protein A)
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                                  </h4>
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                                  <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>
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                                  <img src="https://static.igem.org/mediawiki/2016/6/6c/T--INSA-Lyon--parts7.png" class="img-responsive center-block">
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                              </div>
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                              <div class="subtitle" id="characptac">
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                                  <h2 class="title">Characterization of pTAC promoter</h2>
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                                  <span class="line"></span>
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                                  <p>
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                                    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>.
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                                  </p>
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                                  <h4>
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                                      <a href="http://parts.igem.org/Part:BBa_K1934040">BBa_K1934040</a>: RFP under control of pTAC promoter
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                                  </h4>
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                                  <p>
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                                    This composite part contains the coding sequence of the RFP reporter gene downstream of pTAC and strong RBS sequences.
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                                  </p>
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                                  <img src="https://static.igem.org/mediawiki/2016/e/ee/T--INSA-Lyon--parts1.png" class="img-responsive center-block">
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                                  <h4>
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                                      <a href="http://parts.igem.org/Part:BBa_K1934050">BBa_K1934050</a>: CFP under control of pTAC promoter.
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                                  </h4>
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                                  <p>
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                                      This composite part contains the coding sequence of the CFP reporter gene downstream of pTAC and strong RBS sequences.
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                                  </p>
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                                  <img src="https://static.igem.org/mediawiki/2016/f/fd/T--INSA-Lyon--parts2.png" class="img-responsive center-block">
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Latest revision as of 22:34, 8 November 2016

iGEM : Parts

 
PARTS
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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.



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