Bacterial expression/purification of GFP?
'Mike' Michael J. Moser
moser at U.WASHINGTON.EDU
Mon Sep 16 12:25:37 EST 1996
John
I believe that S65T-GFP can be purified as a 6-His fusion protein. I know
that it expresses like gangbusters from a pRSETB (Invitrogen) construct.
MM
Mike Moser Tel: 206-543-6585
UW Department of Pathology FAX: 206-543-3644
Box 357470 moser at u.washington.edu
Seattle, WA 98195 http://weber.u.washington.edu/~moser
On Sat, 14 Sep 1996, John Fowler wrote:
> Looking for advice/info from others with experience:
>
> I'd like to express GFP as a bacterial fusion protein with either GST or
> MBP, purify it via affinity chromatography, and then cleave off the GFP
> with a site-specific protease. (Using standard bacterial expression
> vectors). The purified (hopefully fluorescent) protein would then be used
> for microinjection.
>
> Has anyone tried this already, and does GFP fluorescence survive the
> purification procedure?
>
> Thanks,
> John
>
> John Fowler
> jfowler at email.unc.edu
>
>
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