Green Fluorescent Protein (GFP): FACS?
cocea at bisance.citi2.fr
Wed May 3 11:55:51 EST 1995
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.
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