I am currently trying to use various GFP vectors to measure drug
induced promotor activity in a CHO cell system.
I have used the same system using constructs of CAT vectors which
have given consistent results, however I am finding the GFP system
fails to give any detectable results, even when using the CMV driven
control vectors.
The vectors I am using are the CytoGEM topaz and sapphire CMV and
Basic vectors from Packard.
I am transiently transfecting them into CHO cells plated in a 96
well plate using Qiagens SuperFect reagent and trying to measure
expression from 24 hours onward.
I am currently unable to detect any difference between fluorescence
from CHO cells transfected and non-transfected.
Can anyone help?
Is there a standard protocol available for this kind of work?
I have spoken to representatives from Pakard in the UK, and they know
of no-one in the UK who is using their plasmids for this kind of
work.
Any information which you may think could help me would be gratefully
appreciated.
Thank you in advance
Ric Anney
PhD student
Department of Psychological Medicine
University of Wales College of Medicine
Heath Park
Cardiff
CF4 4XN
anney at cf.ac.uk
