GST-fusion system question

Seng-Lai Tan sltan at
Sat Mar 1 19:08:14 EST 1997

what type of host cells are you using?  have you tried including some
protease inhibitors in your lysing buffer?  what kind of conditions
(temperature, IPTG concentration, hours of induction, etc.) have you tried
for the purification?

On Thu, 30 Jan 1997, Bart K. Kwieciszewski wrote:

> I am trying to purify GST-NS5A fusion proein, but I'm having extreme
> difficulties getting a a high yield expression.  First the fusion protein
> is unstable, and second, the NS5A protein by itself is very unstable.  I
> would like to purify about 500 micrograms of the full length product from
> 1-2 liters of original culture, but I'm ending up with about 100.  HELP!
> Bart K. Kwieciszewski
> University of Washington
> Regional Primate Research Center
> Department of Microbiology
> HSB H-331

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