Use of GFP as a reporter vector

RICHARD J ANNEY Anney at cardiff.ac.uk
Thu Jul 16 08:52:59 EST 1998


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




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