Expression of GST fusion
pkursula at zombie.oulu.fi
Thu Mar 14 06:17:57 EST 1996
> > Hi, I'm using the GST fusion protein expression system and getting fairly
> > good expression. However, when purifying the protein, a significant amount
> > of the fusion protein is getting broken down, and I get an enormous band
> > of GST+extra a.a. running at ~30kD (GST is 26kD) immediately after cell
> > lysis. I've used a wide array of protease inhibitors which seem to have
> > little effect. Has anyone had any luck in getting rid of this band?
I got rid of the degradation almost completely by reducing the induction
time. 6 hours and the protein was degraded, 2 hours and it was just fine
one nice clean band... you could try reducing your IPTG concentration, too.
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