Guidelines for GST fusion
T. S. Pillay
tpillay at ucsd.edu
Sat Mar 5 02:04:48 EST 1994
Subject: Guidelines for GST fusion
From: Jason Lei, leij at mentor.cc.purdue.edu
Date: Thu, 3 Mar 1994 13:34:36 GMT
In article <CM3CDp.AAp at mentor.cc.purdue.edu> Jason Lei,
leij at mentor.cc.purdue.edu writes:
>One graduate student in our lab had problem in using pGex for making
>fusion protein. She only saw the right size (~64 kD) once; all other
>trials revealed only a size (~50 kD) of seemingly degraded product.
>She wondered if there are general guidelines (or source of such
>guidelines) for making proteins by GST fusion.
>By the way, she tried both DH5a and JM105 (cultured overnight in 2XYT
>with or without IPTG induction) with the same results. She certainly
>would appreciate any ideas on this problem or guidelines for using pGex.
The recommended strain for pGex is DH5a. I presume she has read the
Pharmacia handbook which comes with pGex and provides troubleshooting
guidelines. Every fusion protein is different - that's the guideline I
follow -you have to play around to find the right conditions. I would
suggest using a protease-deficient strain if proteolysis is a problem.
More information about the Methods