Help!! E coli fusion protein expression!
nantel at biotech.lan.nrc.ca
Tue Aug 15 13:35:39 EST 1995
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.
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.
Biotech Research Institute
National Research Council Canada
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