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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