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problem with pGL3basic luc activity

transfection

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2 replies to this topic

#1 mcunaj

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Posted 10 September 2012 - 01:16 AM

I have some problem because the pGL3basic alone (transfected into hela cells) answers to dose dependant assay. I tried with different doses of expression vector for the expression of the cjun protein which repress the basal activity of the pGL3basic.I obtained the same results when I cloned the promoter of my gene. I can't conclude the results of my transfection.I don't know if the repressive effect that I observed with cjun is due to the promoter of my gene or is due to the pGL3basique.
anyone had the same problem with pGL3basique
anyone can help?
thanks

#2 srpres

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Posted 11 September 2012 - 05:15 AM

pGL3 has relatively high background activity which can be inducible upon stimulation with exogenous agents. I think its because of some kind of cryptic promoter activity. I know that pXPG-Luc has lower background activity (like 20 x lower than pGL3 Basic in HEK293 T) but also I strongly suspect the new version pGL4 Basic is better as well. pXPG is avaialble from Peter Cockerill and pGL4 from Promega.

Here is a useful reference:

http://www.ncbi.nlm....=pXPG Cockerill

#3 mcunaj

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Posted 13 September 2012 - 12:49 AM

Thanks for your response . I will try the new version pGL4.





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