I would start by playing with the induction conditions there...time, IPTG
concentration, temperature etc.
On Sun, 12 Dec 1999, Christof Gaenzler wrote:
> Hi,
>> Colin Rasmussen wrote:
> > Alex Blais wrote:
> > > I am trying to purify a recombinant protein fused to GST overproduced in
> > > E.coli. I have some problems during the purification steps, and I
> > > hypothesised that homodimers formation mediated by the GST moiety could
> > > be the cause of the problem.
> >
> > Two questions:
> >
> > How do you know there are dimers?
> >
> > Why does that pose a problem?
>> In a SDS-page, the dimers are no longer stable and one can see C-terminal
> degradation products that are as well purified because of the intact
> N-terminal GST-part. If you want to have a pure full length protein you have
> to do a second purification step.
>> --
> German Cancer Research Center (DKFZ)
> Applied Tumorvirology - ATV F0200
> Christof Gaenzler
> INF 242
> 69120 Heidelberg
> T: +49-6221-42-4937
> F: +49-6221-42-4932
>>
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