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Expression of GFP in eukaryotic cells.

Dave Bates dobates at ucdavis.edu
Mon Jul 17 17:38:27 EST 1995

In article <3u5beh$a9a at bisance.citi2.fr>
villartay at citi2.fr writes:

> I tried to use the pGFP-C1 vector as control in transfection experiments and 
> did not get any expression.
> (using another control showed that the transfection was ok though)
> The list of mammalian GFP failure is getting longer
> jean pierre de Villartay
> villartay at citi2.fr

After over a year of trying various different GFP expression vectors in
BAEC and NIH 3T3 I'm still no closer to getting any fluorescence. I do
get transcription though (RT PCR works fine, but PCR on the RNA gives
nothing), and I can detect it (from a slide from Clontech, and from
bacterial expression) so I don't think it's likely to be a promoter
problem. I would put money on it that it's a problem either with
translation of the RNA to give a protein with a functioning fluorescent
moiety or rate of degradation of normal GFP. It may be a problem with
formation of the fluorophore after normal production of GFP. The most
puzzling thing is why do some people get it to work and others don't?

Dave Bates PhD                                     Tel: 916 752-7081
Postdoctoral Researcher                            Fax: 916 758-2554 
Dept of Human Physiology                         email:
dobates at ucdavis.edu
University of California at Davis                drink: anything
Davis, California 95616, USA            No I don't have a sense of
                                                now buy me a beer      

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