Difference between revisions of "Part:BBa K1065310"

Revision as of 07:59, 23 September 2013

Blue light circuit with inverter for the production of amilCP

This part is a blue light sensing device: it consists of a Blue light sensor with its response regulator (Bba_K592016), the RR dependent promoter (Bba_K592006) and an inverter cassette (cI and pLambda) which are needed to produce the reporter (amilCP) when blue light (470 nm) is present.Dark causes the turning off, thus inhibits the production of the reporter.

Everything is under the control of a constitutive promoter (anderson family).

This part was cloned and successfully characterized by UNITN-Trento 2013 iGEM team in order to test protein transcription and then replace the blue chromoprotein with an ethylene forming enzyme (EFE). The final goal is to design an ethylene producing device that is induced by blue light to control and speed up fruit ripening.

Parts from 2011 Uppsala-Sweden team and 2006 Berkeley team were used.


SAFETY NOTES: this part does not have safety concerns.

Usage and Biology

YF1, the blue light sensor, is a fusion protein of the LOV blue light sensor domain of Bacillus subtilis (YtvA) and FixL histidine kinase domain (from Bradyrhizobium japonicum).
In the dark, the autophosphorylated YF1 phosphorylates FixJ, its Response Regulator, which activates the pFixK2 promoter allowing the expression of the inverter cI. cI instead inhibits pLambda activity thus amilCP transcription.
Under constant illumination with blue light net kinase activity is strongly suppressed, consisting in a consequent inactivation of pFixK2: the outcome is AmilCP production.
we characterized ths part in E. coli using cells NEB10b

CHARACTERIZATION Sequence and Features