Difference between revisions of "Part:BBa K1065309"

Revision as of 20:49, 28 September 2013

light regulated circuit producing amilGFP+EFE at dark

This part includes blue light sensor YF1, its RR FixJ, pFixK2, the reporter amilGFP and EFE: 2-oxoglutarate oxygenase/decarboxylase is an Ethylene Forming Enzyme that catalyzes ethylene biosynthesis from 2-oxoglutarate. This enzyme was initially purified from Pseudomonas siringae pv. phaseolicola PK2, a 2-oxoglutarate-dependent ethylene producing bacterium.

Everything is under the regulation of pLac.
In the presence of blue light (470 nm) the production of the reporter amilGFP and EFE enzyme is inhibited.
In the absence of blue light the device is activated, thus producing amilGFP+EFE.
This part was cloned by UNITN-Trento 2013 iGEM team for the creation of an aerobically engineered pathway for the control of fruit ripening: unlike Bba_k1065311, it doesn't have any inverter cassette, so blue light acts like an inhibitor of the circuit rather than an inducer.

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 amilGFP and EFE. Under constant illumination with blue light net kinase activity is strongly suppressed, consisting in a consequent inactivation of pFixK2.

We characterized only part BBa_K1065311, the blue light dependent circuit producing amilGFP+EFE that contains the inverter cassette.

Safety

Ethylene is a compound that can be explosive at high concentrations (between 2.7% and 34% v,v). We suggest to manage this part carefully. (See BBa_K1065001 for more details.)


Sequence and Features