Team:Cambridge/Bioluminescence/Luciola cruciata

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Latest revision as of 14:55, 21 October 2010

Project Firefly: Luciola cruciata

Photinus pyralis (source)

The Japanese firefly, Luciola cruciata is notable in that nearby fireflies flash in synchrony, making the trees pulsate with green light which fishermen use as a guide to bring them home. The Cambridge team selected Luciola cruciata because [http://www.ncbi.nlm.nih.gov/pubmed/1946326 Kajiyama and Nakano (1991)] had described a suite of single amino acid changes which significantly changed the wavelength of peak emission.

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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:
+The Japanese firefly, ''Luciola cruciata'' is notable in that nearby fireflies flash in synchrony, making the trees pulsate with green light which fishermen use as a guide to bring them home.  The Cambridge team selected Luciola cruciata because [http://www.ncbi.nlm.nih.gov/pubmed/1946326 Kajiyama and Nakano (1991)] had described a suite of single amino acid changes which significantly changed the wavelength of peak emission.
-* [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.
+==Site directed mutagenesis with Gibson Assembly==
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