Difference between revisions of "Part:BBa K1351036"

Revision as of 09:17, 27 October 2014

Quorum sensing two component system (AIP-II sensing histidine kinase agrC, response regulator agrA)

Quorum sensing two component system derived from Staphylococcus aureus N315. This part consists of the AIP-II sensing histidine kinase AgrC and the response regulator AgrA. Upon detection of autoinducing peptide II (AIP-II), AgrC phosphorylates AgrA, which then bindes to distinct binding sites and activates the P2 and P3 promoters. This part should only work for AIP-II. Other AIPs should lead to inhibition.

This part was generated in a modified version of RFC25, where a strong Shine Dalgarno Sequence (SD) is included, and has the following prefix and suffix:

Sites of restriction enzymes generating compatible overhangs have the same color:
EcoRI and PstI in blue, NotI in green, XbaI and SpeI in red, NgoMIV and AgeI in orange. Shine-Dalgarno sequence and stop codons are underlined.

This part is used in the 2014 LMU-Munich iGEM project [http://2014.igem.org/Team:LMU-Munich BaKillus].

Luminescence of B. subtilis P_xylA - agrCA / P2,P3 was measured in MCSE inducing with 0, 0.02, 0.2 % xylose. This plot shows the luminescence readout normalised over OD600 over the course of 6hours. The higher output in xylose induced samples shows the successfull expression of agrCA in B. subtilis.

In the following graph, a dose response curve of the two CSP inducible promoters P_comC and P_comAB is depicted. The dose response curve was generating using luminescence values two hours after inducing with CSP. Sequence and Features