Difference between revisions of "Team:Edinburgh/Composite Part"

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<h2> Composite Parts</h2>
 
  
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<h4>Note</h4>
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<div id="custom-bootstrap-menu" class="navbar navbar-default navbar-fixed-top" role="navigation">
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<p>In order to be considered for the <a href="https://2015.igem.org/Judging/Awards#SpecialPrizes">Best New Composite Part award</a>, you must fill out this page. Please give links to the Registry entries for the Composite parts you have made. Please see the Registry's <a href="http://parts.igem.org/Help:Parts#Basic_and_Composite_Parts"> Help:Parts page</a> for more information on part types.</p>
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    <div class="container-fluid">
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            <button type="button" class="navbar-toggle" data-toggle="collapse" data-target=".navbar-menubuilder"><span class="sr-only">Toggle navigation</span><span class="icon-bar"></span><span class="icon-bar"></span><span class="icon-bar"></span>
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            <ul class="nav navbar-nav navbar-right">
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              <li class="active">
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                  <a href="https://2015.igem.org/Team:Edinburgh">Home</a></li>
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                  <li class="dropdown">
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                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">About<span class="caret"></span></a>
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                    <ul class="dropdown-menu" role="menu">
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Team">Team</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Attributions">Attributions</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Collaborations">Collaboration</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Sponsors">Sponsors</a></li>
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                    </ul>
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                  <li class="dropdown">
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                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">Project<span class="caret"></span></a>
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                    <ul class="dropdown-menu" role="menu">
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Description">Overview</a></li>
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                    <!-- <li><a href="https://2015.igem.org/Team:Edinburgh/Experiments">Experiments</a> </li> -->
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/HeroinBiosensor">Heroin Biosensor</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/DNPBiosensor">DNP Biosensor</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/PMABiosensor">PMA Biosensor</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/CBD">Making it Stick</a></li>           
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Results">Limits of Detection</a></li>
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                    </ul>
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                  </li>
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                  <li class="dropdown">
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                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">Parts<span class="caret"></span></a>
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                    <ul class="dropdown-menu" role="menu">
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Parts">Team Parts</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Basic_Part">Basic Parts</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Composite_Part">Composite Parts</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Part_Collection">Part Collection</a> </li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Improved_Part">Improved Parts</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Characterisation_Part">Improved Characterisation</a></li>
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                    </ul>
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                  </li>
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                  <li class="dropdown">
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                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">Dry Lab<span class="caret"></span></a>
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                    <ul class="dropdown-menu" role="menu">
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Modeling">Modeling</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Design">Design</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Software">Software</a></li>
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                    </ul>
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                  </li>
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                  <li class="dropdown">
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                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">Human Practices<span class="caret"></span></a>
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                    <ul class="dropdown-menu" role="menu">
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Practices">Our Story</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Safety">Safety</a> </li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Legality">Legality</a> </li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Outreach">Outreach</a> </li>
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                    </ul>
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                  </li>
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                  <li class="dropdown">
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                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">InterLab<span class="caret"></span></a>
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                    <ul class="dropdown-menu" role="menu">
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/InterLab">InterLab</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/InterLabBook">LabBook</a> </li>
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                    </ul>
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                  </li>
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                  <li class="dropdown">
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                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">Notebook<span class="caret"></span></a>
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                    <ul class="dropdown-menu" role="menu">
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Project/Protocols">Protocols</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Notebook/HeroinPurity">Heroin Purity</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Notebook/PMADetection">PMA Detection</a> </li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Notebook/DNPDetection">DNP Detection</a> </li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Notebook/FluidDynamics">Fluid Dynamics</a> </li>
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                    </ul>
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                  </li>
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                  <li><a href="https://2015.igem.org/Team:Edinburgh/MedalCriteria">Accomplishments</a></li> 
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            </ul>
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        </div>
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    </div>
 
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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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          <header class="intro">
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            <div class="row">
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              <div class="col-md-12">
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                <h1 class="brand-heading">Composite Parts</h1>
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                <p class="intro-text">
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                </p>
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                <div align="center">
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                    <a href="#cd-team">
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                        <span class="arrowtext">Scroll down to read more</span>
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                        <img src="https://static.igem.org/mediawiki/2014/3/3e/Aalto_Helsinki_Nuoli.png" class="arrow">
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            <div class="row">
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                <div class="col-lg-12 text-center">
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                    <h2 class="section-heading">mRFP</h2>                   
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            <div class="panel-heading">
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                <h4 class="panel-title">
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne">mRFP driven by LacI promoter (BBa_K1615090)</a>
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                </h4>
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                    <p>mRFP (BBa_K1615089) driven by the IPTG inducible PLlac 0-1 promoter (BBa_K314103). This allows the <i>mRFP</i> gene to express mRFP when induced with IPTG.
 
</p>
 
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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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            <div class="panel-heading">
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                <h4 class="panel-title">
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseTwo">CBDcenA+Linker fused to mRFP in RFC25 (BBa_K1615091)</a>
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                </h4>
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                    <p>mRFP (BBa_K1615089) fused to the C terminal of CBDcenA + C-terminal linker (BBa_K1321339) in RFC25. If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.
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<br>
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<img src="https://static.igem.org/mediawiki/2015/2/20/CBD_RFP_folding.jpeg">
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<br>
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To test the RFP folding when fused to a CBD we placed 200µL of crude cell lysate in a 96 well plate and observed the fluorescence under a blue light transilluminator.
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</p>
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            <div class="panel-heading">
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                <h4 class="panel-title">
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseFour">mRFP fused to CBDcenA+Linker in RFC25 (BBa_K16150092)</a>
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                </h4>
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            <div id="collapseFour" class="panel-collapse collapse">
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                <div class="panel-body">
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                    <p>mRFP (BBa_K1615089) fused to the N terminal of CBDcenA + C terminal linker (BBa_K1321339) in RFC25. If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.
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<br>
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<br>
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<img src="https://static.igem.org/mediawiki/2015/2/20/CBD_RFP_folding.jpeg">
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<br>
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<br>
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We tested the dissociation of this part on Whatman 54 using isostandard chads. This involved incubating the chads in the crude cell lysate for 10 minutes and then washing them with PBS for various periods time to test the binding affinity of this CBD.
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<br>
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<br>
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We tested the dissociation of this part on Whatman 54 using isostandard chads. This involved incubating the chads in the crude cell lysate for 10 minutes and then washing them with PBS for various periods time to test the binding affinity of this CBD.
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</p>
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        <div class="panel panel-default">
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            <div class="panel-heading">
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                <h4 class="panel-title">
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseFive">Linker +dCBD fused to mRFP in RFC25 (BBa_K1615093)
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</a>
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                </h4>
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            </div>
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            <div id="collapseFive" class="panel-collapse collapse">
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                <div class="panel-body">
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                    <p>mRFP (BBa_K1615089) fused to the C terminal of dCBD + N terminal linker (BBa_K1321340). If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.
 +
<br>
 +
<br>
 +
<img src="https://static.igem.org/mediawiki/2015/2/20/CBD_RFP_folding.jpeg">
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<br>
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To test the RFP folding when fused to a CBD we placed 200µL of crude cell lysate in a 96 well plate and observed the fluorescence under a blue light transilluminator.
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Latest revision as of 19:10, 20 November 2015

Composite Parts

mRFP

mRFP driven by LacI promoter (BBa_K1615090)

mRFP (BBa_K1615089) driven by the IPTG inducible PLlac 0-1 promoter (BBa_K314103). This allows the mRFP gene to express mRFP when induced with IPTG.

CBDcenA+Linker fused to mRFP in RFC25 (BBa_K1615091)

mRFP (BBa_K1615089) fused to the C terminal of CBDcenA + C-terminal linker (BBa_K1321339) in RFC25. If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.

To test the RFP folding when fused to a CBD we placed 200µL of crude cell lysate in a 96 well plate and observed the fluorescence under a blue light transilluminator.

mRFP fused to CBDcenA+Linker in RFC25 (BBa_K16150092)

mRFP (BBa_K1615089) fused to the N terminal of CBDcenA + C terminal linker (BBa_K1321339) in RFC25. If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.



We tested the dissociation of this part on Whatman 54 using isostandard chads. This involved incubating the chads in the crude cell lysate for 10 minutes and then washing them with PBS for various periods time to test the binding affinity of this CBD.

We tested the dissociation of this part on Whatman 54 using isostandard chads. This involved incubating the chads in the crude cell lysate for 10 minutes and then washing them with PBS for various periods time to test the binding affinity of this CBD.

Linker +dCBD fused to mRFP in RFC25 (BBa_K1615093)

mRFP (BBa_K1615089) fused to the C terminal of dCBD + N terminal linker (BBa_K1321340). If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.


To test the RFP folding when fused to a CBD we placed 200µL of crude cell lysate in a 96 well plate and observed the fluorescence under a blue light transilluminator.