Team:Cambridge/Bioluminescence/Photinus pyralis

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{{:Team:Cambridge/Templates/headerbar|colour=#96d446|title=Bioluminescence: Firefly Luciferases}}
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{{:Team:Cambridge/Templates/headerbar|colour=#96d446|linkcolour=#557A25|title=Project Firefly: Photinus pyralis}}
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<img src="https://static.igem.org/mediawiki/2010/thumb/f/f2/Firefly.jpg/300px-Firefly.jpg">
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<img src="https://static.igem.org/mediawiki/2010/8/83/Cam-Petri.jpg" style="margin-bottom:10px;">
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<strong>Artist's impression</strong> of what we would like<br /> to achieve
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<i>Photinus pyralis (<a href="http://www.flickr.com/photos/artfarmer/198487523/">source</a>)</i>
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We are investigating a number of firefly luciferases.  As a comparison for our future experiments we will begin by cloning the existing Biobrick for [http://partsregistry.org/Part:BBa_I712019 wild type <i>P. pyralis</i> luciferase] into our strains of interest.
 
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Our project will focus on characterising both this existing luciferase and those we hope to submit to the registry:
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The luciferase of the North American firefly, ''P. pyralis'', is a tried and tested mechanism for creating bioluminescence.  We were aware that a luciferase from this organism was already present in the registry ([http://partsregistry.org/Part:BBa_I712019 BBa_I712019]).  We wanted to improve on this by three techniques:
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* [https://2010.igem.org/Team:Cambridge/Codons Codon optimisation] for expression in E. coli to increase the rate of translation
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* Using a mutant with increased substrate affinity
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* Parallel use of the Photinus pyralis [https://2010.igem.org/Team:Cambridge/Bioluminescence/Luciferin_Regeneration luciferin regenerating enzyme] to both relieve inhibition by oxyluciferin and increase availability of luciferin.
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* An enhanced <i>P. pyralis</i> luciferase with increased brightness
 
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* A number of differently coloured versions of <i>L. cruciata</i> luciferase
 
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We intend to synthesise these luciferases on operons also containing the LRE for the firefly species in question, and then to quantify a PoPS to photons function.
 
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Latest revision as of 09:46, 18 October 2010

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