2 replies to this topic

[mcunaj]

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

[srpres]

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

[mcunaj]

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