Revision as of 04:35, 27 October 2010

BioBricks

The Cambridge iGEM team has submitted 20 parts to the Registry of Standard Biological Parts. Many of these parts have been extensively characterised. This data is stored in the Parts Registry, you can click on any BioBrick to view its technical datasheet.

Featured parts

BBa_K325219

Red luciferase and LRE operon from L. cruciata under pBAD

This part generates the luciferase and luciferin regenerating enzyme proteins from the Japanese firefly, Luciola cruciata, when induced by L-arabinose. The luciferase produced has a point mutation Ser286Asn which gives it a redshifted emission spectrum as compared to wild-type. The sequence of nucleotides is codon optimised for expression in E. coli.

BBa_K325909

V. fischeri luxCDABE under pBAD

This part generates luxCDABE proteins from the bioluminescent bacterium, Vibrio fischeri, when induced by L-arabinose. The BioBrick produces light from basic metabolites found in E. coli, luxCDE produce the substrate used by luxAB for bioluminescence. The part creates a blue light.

Parts based on bacterial systems

BBa_K325902	V. fischeri luxCD This is a translational unit designed to produce V. fischeri luxC and luxD genes when placed under a suitable promoter. The DNA sequence is codon optimised for expression in E. coli. Both proteins are part of the fatty aldehyde synthesis pathway. luxC is a reductase and luxD is a acyltransferase. LuxE must also be supplied to complete the pathway for fatty aldehyde synthesis. Such fatty aldehydes are the substrate for the luxAB bacterial luciferase.
BBa_K325903	V. fischeri luxEG This is a translational unit designed to produce V. fischeri luxE and luxG genes when placed under a suitable promoter. The DNA sequence is codon optimised for expression in E. coli. luxE functions as the synthetase creating fatty aldehydes for bioluminescence. LuxG is a flavin reductase.
BBa_K325905	luxCDEG pLambda AB This part constitutively expresses the Xenorhabdus luminescens luxAB luciferase already in the registry. It also features a translational unit for the substrate generating luxCDEG from Vibrio fischeri. This means that only when supplied with PoPs will it produce light output.
BBa_K325906	pBAD luxCDEG pLambda AB This is the same as the part above but was built for testing induction by adding the pBAD promoter, making light inducible by addition of L-arabinose.

Firefly luciferases

BBa_K325101	Luciferase from Photinus pyralis with increased substrate affinity This is a translational unit for a mutated and codon-optimised form of the luciferase from the North American firefly, Photinus pyralis. It is described in Fujii et al. 2007 as having a 10 times higher substrate affinity and luminescence output compared to wildtype.
BBa_K325108	P.pyralis luciferase under pBAD This is the same as the part above but placed under the pBAD promoter induced by L-arabinose for characterisation.
BBa_K325211	L. cruciata luciferase (red mutant) This is a translational unit for a mutant (Ser286Asn) and codon-optimised luciferase from the Japanese firefly, Luciola cruciata. It is deeply red-shifted as compared to wild type.
BBa_K325218	L. cruciata luciferase under pBAD This is the same as the part above but placed under the pBAD promoter induced by L-arabinose for characterisation.

Firefly luciferin regenerating enzymes

BBa_K325102	P. pyralis Luciferin Regenerating Enzyme This is a translational unit for a codon-optimised Luciferin Regenerating Enzyme (LRE) from the North American firefly, Photinus pyralis.
BBa_K325202	L. cruciata Luciferin Regenerating Enzyme This is a translational unit for a codon-optimised Luciferin Regenerating Enzyme (LRE) from the Japanese firefly, Luciola cruciata.

Firefly luciferase, LRE operons

BBa_K325210	Luciola cruciata LRE and luciferase This translational unit combines the luciferin regenerating enzyme (BBa_K325202) and red-shifted luciferase (BBa_K325211) from the Japanese firefly, Luciola cruciata. The luciferin regenerating enzyme helps to strengthen and sustain light output.
BBa_K325100	Photinus pyralis LRE and luciferase This part combines the luciferin regenerating enzyme (BBa_K325102) and luciferase (BBa_K325101) from the North American firefly, Photinus pyralis. The luciferin regenerating enzyme helps to strengthen and sustain light output.
BBa_K325109	Photinus pyralis LRE and luciferase under pBAD This part is the part above expressed under pBAD for characterisation.
BBa_K325209	L. cruciata LRE and luciferase (wildtype) under pBAD This part combines the Luciferin Regenerating Enzyme and luciferase from the Japanese firefly, Luciola cruciata. It is expressed under pBAD.
BBa_K325229	L. cruciata LRE and luciferase (green) under pBAD This is the same as the part above but with a Val239Ile mutation which green-shifts its emission spectrum.
BBa_K325239	L. cruciata LRE and luciferase (red) under pBAD This is the same as the part above but instead has a Gly326Ser mutation which red-shifts its emission spectrum.
BBa_K325249	L. cruciata LRE and luciferase (scarlet) under pBAD This is the same as the part above but instead has a His433Tyr mutation which red-shifts its emission spectrum.
BBa_K325259	L. cruciata LRE and luciferase (yellow) under pBAD This is the same as the part above but instead has a Pro452Ser mutation which shifts its emission spectrum to peak at a yellow wavelength.

Summary table

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From 2010.igem.org