GST dimers

Petri Kursula pkursula at cc.oulu.fi
Tue Dec 14 09:10:19 EST 1999


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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Petri Kursula           "I am somehow less interested in the weight and
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Petri.Kursula at oulu.fi  certainty that people of equal talent have lived
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http://cc.oulu.fi/~pkursula                        -- Stephen Jay Gould 
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