I am going to boldly attempt the purification of a viral membrane protein and want to try several expression systems to see what works best.
One I would like to try is pBAD because (in my hands with previous proteins), the AraBAD promoter/AraC repressor system gives excellent transcriptional control - including complete repression prior to induction, which seems hard to achieve in T7 systems. But this will be my first membrane protein, and I'm told NEB 10-beta may not be the best cell line for toxic membrane proteins. Does anyone know another strain with the arabinose metabolism mutations (araBADC- or araD139 Δ( araleu)7697) that's good for membrane proteins?
Hi,
pBAD is good one to try for membrane proteins (if you're sticking to E. coli). With regards to strains I've had good experiences with membrane protein expression in pBAD vectors using MC1061 and TOP10 cells.
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