Promoter activity

Theodosios Kyriakou theodosios73 at yahoo.com
Wed Feb 16 07:14:17 EST 2000


Hi all
This is the problem that is keeping me up at nights and I hope some of
you will help me solve it.
I am transfecting plasmid constructs in Hek 293 cells using a liposome
based method. My plasmids contain a reporter gene(CAT) with my promoter
inserted right in front of it.When I am assaying for CAT activity I get
really high activity from BOTH my "insertless" control plasmid and the
plasmid with the promoter insert.Normally one would expect only minimal
transcription from the control plasmid but this obviously is not the
case here. I tried to normalise my results co-trasfecting with a pRSv
beta gal plasmid but my cells don't seem to like it very much.

I would appreciate if someone can think of something.
Cheers

--

Theodosios Kyriakou
Biological Sciences,WarwickUniversity ,CV4 7AL
tel:++44-1203-522440,++44-1203-572572 ext.30164


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