Expression of GST fusion

Petri Kursula 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.

Pete




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