pGL3 / pGL2 dilemma: anybody working with these ?

Alt Zantema A.ZANTEMA at BIOCHEMISTRY.MedFac.LeidenUniv.NL
Tue Feb 11 15:53:22 EST 1997


un691cs at genius.embnet.dkfz-heidelberg.de (Clemens Suter-Crazzolara)
wrote:

>Hello, 

>we have cloned our potential promoter in pGL3, and made all the deletion-
>studies. Now we have a problem: in certain cell lines, we find that our
>constructs are much less active than the pGL3 vector.
>This is difficult to publish.
>I read on this group that pGL2 has lower background levels than pGL3;
>but to reclone my 25 fragments and repeat all the transfections ands
>assays does not sound very atractive.

>Could any one give me advice how to proceed ? is there perhaps a publication
>where pGL2 is compared to pGL3, or which states that pGL3 results in rela-
>tively high BG-levels, which I could refer to ?

>thanks, 

>clemens Suter-Crazzolara    (answers to my e-mail address would be appreciated)
>University of Heidelberg
>Dept. Anatomy and Cellbiologie

As far as I know, pGL3 (and also pGL2) is a construct with a
luciferase with no promoter in front of it (only a cloning site). So
for a control at least a minimal promoter should be introduced. It is
unclear why the background of pGL3 is higher then pGL2. The main
difference is that in pGL3 crytic AP-1/ATF sites are removed.
Therefore, for studies of promoters in which AP-1 and/or ATF are
involved certainly pGL3 is recommended.

Alt Zantema
A.ZANTEMA at BIOCHEMISTRY.MedFac.LeidenUniv.nl




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