Is a lac promoter with a CRP binding site active in pseudomonas syringae DC3000?

10 October 2013 0 6K Report

I cloned pseudomonas genes downstream to a lac promoter in pUCP24 and pBBR1-MCS2. I am trying to do a western blot and not able to see any bands. VRF, a CRP ortholog is present in pseudomonas syringae DC300 but it is not well characterized yet. CRP's conformation changes when it interacts with cAMP and binds upstream of lac promoter causing DNA bending. This will facilitate RNA pol binding which initiates transcription. I am assuming VRF doesnt bind at CRP binding site and hence lac promoter is inactive in pseudomonas. Am i right?

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