Team:Cambridge/Bioluminescence/Photinus pyralis

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Revision as of 09:35, 18 October 2010

Project Firefly: Photinus pyralis

Photinus pyralis (source)

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:

Codon optimisation for expression in E. coli to increase the rate of translation
Using a mutant with increased substrate affinity
Parallel use of the Photinus pyralis luciferin regenerating enzyme to both relieve inhibition by oxyluciferin and increase availability of luciferin.

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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.
+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:
+* [https://2010.igem.org/Team:Cambridge/Codons Codon optimisation] for expression in E. coli to increase the rate of translation
+* Using a mutant with increased substrate affinity
+* 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.
-Our project will focus on characterising both this existing luciferase and those we hope to submit to the registry:
-* An enhanced <i>P. pyralis</i> luciferase with increased brightness
-* A number of differently coloured versions of <i>L. cruciata</i> luciferase
-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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