Green Fluorescent Protein (GFP): FACS?

Cocea cocea at bisance.citi2.fr
Wed May 3 11:55:51 EST 1995


Hello!

The use of the Green Fluorescent Protein to evaluate the transcription rate
from a promoter seems an interesting application. We are trying to make a
construct containing GFP and several regulatory sequences for a given gene in
order to transfect this construct into cells that produce factors that interact
with the mentioned regulatory sequences.

I know there have been at least two reports on the use of a fluorescence 
activated cell sorter with cells (COS-1 and HEK293) transfected with a vector
exprtessing GFP and that these reports have been mailed to this newsgroup 
(Peter (Univ of Pennsylvania) and Nick Vercammen (Belgium)). However, I only
got this information when it was too late. Therefore I would be much obliged
to any person who might have information on this subject. We could thereafter
exchange information on this subject. I currently use standard CAT-assays to
evaluate the transcription for a promoter/silencer couple but I think the use
of a FACS might help optimize this method a little bit, thai is I hope GFP
detection is sensitive enough to allow quantification of the synthesized
protein after 40-44 hrs of transient transfection.

Some people from Germany told us that they had trouble maintaining GFP-gene
transfected cells in culture on dishes that were not made of glass. Do you
have any information on that?

Thank you very much for your help.

Laurentiu COCEA
INSERM U373
Institut Necker
156, rue de Vaugirard
75730 Paris Cedex 15
France
tel: +33-1-40 61 53 84
fax: +33-1-40 61 55 90 (or +33-1-40 61 56 82 (10:00-17:00 GMT))
internet: cocea at citi2.fr




More information about the Methods mailing list