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?


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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
humour 
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