Cloning and Expression vectors - Which is the best Expression vector to use? (Oct/02/2006 )
I need to clone a cDNA into an expression vector. The purpose is to make as much of the polypeptide as possible so as to send it off to get an antibody raised against it.
I would like to know what vector you would recommend for this, with the caviat that I want nothing to do with Topoisomerase what so ever. It would be nice if this vector offered some sort of positive selection, like blue/white screening, but I'll even accept that as an uncessesary luxury.
I'm just looking for a vector that will express my gene very highly, and have some way like a hisXtag so I can purify it and send the protein off, with nothing to do with topoisomerase. Does anyone have any suggestions?
pcDNA3.1 with myc-His , from invitrogen has Histag for purification.
Would suggest that you use the ones already in use by members of your lab, and especially those that encountered no problem using the vector. At least if problems arise, you have people to share the troubleshooting with .
I am also have the main aim as you as in to get as many protein as possible.
Hmm.. the expression vector i am using is the pQE vector from Qiagen. Not too sure whether it will be a good one. It has strong promoter as well as stop codons at the end. Hopefully it will be the great expression vector.