Difference between revisions of "Team:Edinburgh/Composite Part"
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| − | + | <!-- menu --> | |
<div id="custom-bootstrap-menu" class="navbar navbar-default navbar-fixed-top" role="navigation"> | <div id="custom-bootstrap-menu" class="navbar navbar-default navbar-fixed-top" role="navigation"> | ||
<div class="container-fluid"> | <div class="container-fluid"> | ||
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<li><a href="https://2015.igem.org/Team:Edinburgh/DNPBiosensor">DNP Biosensor</a></li> | <li><a href="https://2015.igem.org/Team:Edinburgh/DNPBiosensor">DNP Biosensor</a></li> | ||
<li><a href="https://2015.igem.org/Team:Edinburgh/PMABiosensor">PMA Biosensor</a></li> | <li><a href="https://2015.igem.org/Team:Edinburgh/PMABiosensor">PMA Biosensor</a></li> | ||
| − | <li><a href="https://2015.igem.org/Team:Edinburgh/CBD">Making it Stick</a></li> | + | <li><a href="https://2015.igem.org/Team:Edinburgh/CBD">Making it Stick</a></li> |
| − | <li><a href="https://2015.igem.org/Team:Edinburgh/Results"> | + | <li><a href="https://2015.igem.org/Team:Edinburgh/Results">Limits of Detection</a></li> |
</ul> | </ul> | ||
</li> | </li> | ||
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<a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">Parts<span class="caret"></span></a> | <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">Parts<span class="caret"></span></a> | ||
<ul class="dropdown-menu" role="menu"> | <ul class="dropdown-menu" role="menu"> | ||
| − | + | <li><a href="https://2015.igem.org/Team:Edinburgh/Parts">Team Parts</a></li> | |
<li><a href="https://2015.igem.org/Team:Edinburgh/Basic_Part">Basic Parts</a></li> | <li><a href="https://2015.igem.org/Team:Edinburgh/Basic_Part">Basic Parts</a></li> | ||
<li><a href="https://2015.igem.org/Team:Edinburgh/Composite_Part">Composite Parts</a></li> | <li><a href="https://2015.igem.org/Team:Edinburgh/Composite_Part">Composite Parts</a></li> | ||
| − | + | <li><a href="https://2015.igem.org/Team:Edinburgh/Part_Collection">Part Collection</a> </li> | |
<li><a href="https://2015.igem.org/Team:Edinburgh/Improved_Part">Improved Parts</a></li> | <li><a href="https://2015.igem.org/Team:Edinburgh/Improved_Part">Improved Parts</a></li> | ||
<li><a href="https://2015.igem.org/Team:Edinburgh/Characterisation_Part">Improved Characterisation</a></li> | <li><a href="https://2015.igem.org/Team:Edinburgh/Characterisation_Part">Improved Characterisation</a></li> | ||
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</ul> | </ul> | ||
</li> | </li> | ||
| − | <li><a href="https://2015.igem.org/Team:Edinburgh/MedalCriteria"> | + | <li><a href="https://2015.igem.org/Team:Edinburgh/MedalCriteria">Accomplishments</a></li> |
</ul> | </ul> | ||
</div> | </div> | ||
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<p class="intro-text"> | <p class="intro-text"> | ||
</p> | </p> | ||
| − | <a href="#cd-team"> | + | <div align="center"> |
| − | + | <a href="#cd-team"> | |
| − | + | <span class="arrowtext">Scroll down to read more</span> | |
| − | + | <img src="https://static.igem.org/mediawiki/2014/3/3e/Aalto_Helsinki_Nuoli.png" class="arrow"> | |
| + | </a> | ||
| + | </div> | ||
</div> | </div> | ||
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<div class="row"> | <div class="row"> | ||
<div class="col-lg-12 text-center"> | <div class="col-lg-12 text-center"> | ||
| − | <h2 class="section-heading"> | + | <h2 class="section-heading">mRFP</h2> |
</div> | </div> | ||
</div> | </div> | ||
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<h4 class="panel-title"> | <h4 class="panel-title"> | ||
| − | <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne"> | + | <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne">mRFP driven by LacI promoter (BBa_K1615090)</a> |
</h4> | </h4> | ||
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<div class="panel-body"> | <div class="panel-body"> | ||
| − | <p> | + | <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> | ||
</div> | </div> | ||
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<h4 class="panel-title"> | <h4 class="panel-title"> | ||
| − | <a data-toggle="collapse" data-parent="#accordion" href="#collapseTwo"> | + | <a data-toggle="collapse" data-parent="#accordion" href="#collapseTwo">CBDcenA+Linker fused to mRFP in RFC25 (BBa_K1615091)</a> |
</h4> | </h4> | ||
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</div> | </div> | ||
| − | <div id="collapseTwo" class="panel-collapse collapse | + | <div id="collapseTwo" class="panel-collapse collapse"> |
<div class="panel-body"> | <div class="panel-body"> | ||
| − | <p> | + | <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. |
| + | <br> | ||
| + | <img src="https://static.igem.org/mediawiki/2015/2/20/CBD_RFP_folding.jpeg"> | ||
| + | <br> | ||
| + | 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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<h4 class="panel-title"> | <h4 class="panel-title"> | ||
| − | <a data-toggle="collapse" data-parent="#accordion" href="# | + | <a data-toggle="collapse" data-parent="#accordion" href="#collapseFour">mRFP fused to CBDcenA+Linker in RFC25 (BBa_K16150092)</a> |
</h4> | </h4> | ||
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</div> | </div> | ||
| − | <div id=" | + | <div id="collapseFour" class="panel-collapse collapse"> |
<div class="panel-body"> | <div class="panel-body"> | ||
| − | <p> | + | <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. |
| − | + | <br> | |
| − | + | <br> | |
| − | + | <img src="https://static.igem.org/mediawiki/2015/2/20/CBD_RFP_folding.jpeg"> | |
| − | + | <br> | |
| − | + | <br> | |
| − | + | 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. | |
| − | + | <br> | |
| − | + | <br> | |
| − | + | 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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</div> | </div> | ||
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<h4 class="panel-title"> | <h4 class="panel-title"> | ||
| − | <a data-toggle="collapse" data-parent="#accordion" href="# | + | <a data-toggle="collapse" data-parent="#accordion" href="#collapseFive">Linker +dCBD fused to mRFP in RFC25 (BBa_K1615093) |
| + | </a> | ||
</h4> | </h4> | ||
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</div> | </div> | ||
| − | <div id=" | + | <div id="collapseFive" class="panel-collapse collapse"> |
<div class="panel-body"> | <div class="panel-body"> | ||
| − | <p> | + | <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"> | ||
| + | <br> | ||
| + | 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. | ||
| + | </p> | ||
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Latest revision as of 19:10, 20 November 2015
mRFP
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.
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 (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.
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.