I am trying to express bacterial efflux pumps in E.coli. The vector I am using for expression is TOPO TA pBAD expression system. When it comes to expression, everything is going fine but during the functional analysis, but in terms of MICS and drug transport assays it becomes difficult to measure the relative degree of expression. I am using E.coli LMG194. Literature suggests using a strain with deleted AcrB would be better. So, which strain I can use for expressional studies (MIC, drug transport assays)? Can I use E.coli JW0451 for expression (MIC).
Yes. An E. coli mutant lacking acrB is very useful as a host to chracterize multidrug efflux pumps.
But my concern is JW0451 is a strain having knocked down AcrB but I couldnt get any info that does it contain promoter required for expression as pBAD Topo system/ can suggest any strain for the same.
Like LMG194,JW0451 is a mutant strain lacking ara genes.
Try to use it and we see how it works.
If you are trying to knock out the indigenous E. coli efflux pumps the best mutant would be a tolC mutant (i.e. TolC is required for most if not all E. coli pumps).
Hello Priyabrata,
Could you please kindly let me know which paper suggested using a strain with deleted AcrB would be better to express efflux pump.
Thank you
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