mark.bowen at stanford.edu
Fri Mar 23 05:00:03 EST 2001
have you looked into trying to lower you expression levels. with some of my
proteins when the expression level gets too high they can aggregate and
become insoluble. perhaps shorter expression times, lower induction levels
or reduced temperature can help keep some of the material soluble.
nadia pantic wrote:
> Hi there folks,
> I need to produce a soluble native recombinant protein. At the moment
> its expressed in
> pET30a and is totally insoluble (the native protein is v. soluble). I've
> tried refolding but the protein has 3 di-sulphide bridges and refolds in
> many different forms (the multiple folded forms have been noticed by
> other groups).
> I want to try to get it soluble by expressing it in a vector that
> targets it to the perplasmic space so I would be very grateful if
> there's any one who has experience with these types of vectors (such as
> pET22b) could give me some advice as in what is a good vector to use,
> wether the manufacturer's claims are exaggerated about solubility etc.
> Thanks Nadia
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