SV: (no subject)

Dag Rune Gjellesvik dagrg at online.no
Sun Mar 25 12:52:22 EST 2001


Hi,
You may try eukaryotic expression if it is really important to get the
protein. We had a small, highly crosslinked protein that was successfully
expressed secreted from Pichia pastoris (yeast). In E. coli it was
impossible.

A bit more work, but better than trying system after system with no result.

Dag Rune


nadia pantic <nadia.pantic at zoology.oxford.ac.uk> skrev i
meldingsnyheter:3ABB4A52.8C2C3A75 at zoo.ox.ac.uk...
> 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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