pGL3 / pGL2 dilemma: anybody working with these ?

Dr. J. Miano jmiano at post.its.mcw.edu
Sat Feb 1 02:41:56 EST 1997


Yeah I know what you mean.  I posted a note a few months back about the
bkgd problem with pGL3; there definitely is higher pGL3 basic luc activity
than its pGL2 cousin, but this seems to be cell type-dependent.

I had a similar problem with a minimal promoter construct cloned into pGL3.
 The activity was two orders of magnitude less than pGL3 basic.  I
published this and speculated about a possible silencer but guess what? I
am wrong.  Although the minimal promoter construct was sequenced multiple
times on both strands no mutations were evident.  My only conclusion (have
not pursued it) is that somehow a point mutation arose in the luc gene
itself because I have since recloned this frag into a new pGL3 and its
fine.  

All I can say is that Promega is not totally correct when they say that the
pGL3 was engineered to reduce background;  someone over there really needs
to explore this issue.

Good luck
-- 
The Love you take is equal to the Love you make--P. McCartney



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