GST fuisons - insoluble -- HELP!!!!!!

News Reader broxholme at
Wed Jul 5 13:10:36 EST 1995


In article <gaw1+-2106951444360001 at 136.142.54.17>, gaw1+ at pitt.edu (Gary
Walker) wrote:

> In article <3s3s1k$qe5 at griffin.ccc.nottingham.ac.uk>, Ian Goodfellow
> <plxigpg at pln1.nottingham.ac.uk> wrote:
> 
> > Dear All
> > 
> > I am having quite a reall 'nightmare' trying to purify a GST fusion 
> > protein.
> How about trying urea and/or DTT with later renaturation of the protein
> for affinity chromatography.  Good Luck.

If you want refernces on using Sarkosyl, lowering growth temperatures and
IPTG concentrations, stressing the cells osmotically during growth (the
betaine-sorbitol method) or some refolding strategies, let me know.  My
main advice is how about changing expression system?  I changed expression
system (from hexahisitidine fusion to GST fusion) and my domain boundaries
(putative only) and had a dramatic improvement in solubility.

Chris



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