Protocol Online logo
Top : Forum Archives: : Protein and Proteomics

GST fusion protein expression problem - (Oct/17/2007 )

Does anyone have experience with this?
I have tried to express my protein which is a glycosyltransferase fused with GST (pGEX 4T-1).And its MW is 43kd() It can be induced in 1ml LB, but can not be induced in a large scale (100ml LB). My bacteria strain is BL21 and my induction condition: 37 deg, 200RPM, 1mM IPTG, induction 12hr.
Can someone give me an idea how to solve this problem? Thanks

-LNY-

You mean you can get your proetin in small scale preparation but not in large one?
this is strange because usually if you can get in a small scale you can get it also in large scale.....but i noticed something you said you induce for 12 hours in 37 degree?
i think its too long usually 3 -4 hours in 37 degree is enough if you want longer period reduce the temperature of incubation.
may this help you

-spanishflower-