Difference between revisions of "Part:BBa K737067"

Revision as of 12:00, 26 September 2012

pLux OmpR hybrid promoter

In order to optimize the function of the lux promoter, We creat a hybrid promoter which combine plux with OmpR, and in this case, plux can be lead to expression by AHL and obtain by phosphorylation of OmpR protein. According to the research of it before, we camp up with a program . Keep the sequencing of Lux Box, -35box and -10box. Then put the sequencing of C3 point of OmpR promoter into -35box and -10box.

Figure structural representation of Plux promoter

Figure : the analysis of conserved sequence of pLux by Weblogo

Plux promoter is organized from the promoter the shining gene of V. fischeri(R0062). Through analyzing common hybrid promoter of Lux by Weblogo , we found that the sequence of Lux Box is conserved. In benefit of LuxR Binding Site, -35box and -10box of the promoter the shining gene of V. fischeri(R0062).

Figure : Our design of pLux-OmpR hybrid promoter

We link the hybrid promoter to the test circuit. For control group experiment, we put strain A to LB medium adding 20% Sucrose solution and not adding any sucrose solution to cultivate in 37% shake flask , and detect the change of OD value and fluorescence value between 12h.

Though high concentration sucrose solution restrains the grain of E.coli, it doesn’t have effect on the work of circuit. In conclusion, we find that high osmotic pressure has no effect on quorum sensing circuit and the activated of plux promoter. Strain A could serve as control group of pLux-OmpR hybrid promoter.

Sequence and Features