Maltose Binding Fusion Protein MBP - which strain for expression? (Oct/03/2006 )
I would like to know if anyone of you has experience with the pMalc2 expression vector of NEB? Is it possible to transform/express the fusion protein in a BL21(DE3) strain? To my knowledge IPTG will produce the T7 polymerase in BL21(DE3), which takes over completely transcription. Is there a chance at all for expression of the MBP fusion protein which is driven by the Tac promotor?
thanks a lot
I have used the pMALp2x. I use regular BL-21 DE3 cells and induce with IPTG. It works fine, but it does leak some and intracellular MBP can reduce your binding yields. The HB101 cells are susposed to reduce cellular MBP, but I have not used them. I also make my own amylose resin, I can give you the protocol if you want. It's easy and cheap.