Help!! E coli fusion protein expression!

[andre]

In article <1995Aug11.155621.7944 at mcrcr6>
huange01 at mcrcr6.med.nyu.edu writes:

> I have tried to express some proteins in pGex2T in Ecoli.  When I
> expressed them in small prep (50 mL), there was some breakdown but not
> significant.  However, when I expressed them in a larger volume
> (300mL), there was a lot of breakdown.  I expressed them at 37
> degree for only an hour after growing them to higher density.  Is
> there any chang that I can do to reduce breakdown of my protein?  It
> is really frustrating.
> Thanks

You may want to try a protease-deficient strain. We observed some major
improvements with UT5600 which is deficient in a membrane-bound
protease that acts during cell lysis.

Good luck,

Andre Nantel
Biotech Research Institute
National Research Council Canada
Montreal, Quebec