Promoter activity

A.F. Simpson AFS7 at
Wed Feb 16 18:41:11 EST 2000

Theodosios Kyriakou wrote:

Hi, Theo!

> 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.

Which plasmid is 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.

Firstly, have you tried a CAT assay on mock transfected 293s (no plasmid
at all, just lipid and the same treatment as the trasnfected cells),
just to make sure it's not the assay step which is giving the weird

Secondly, which 293 cells are you using?  As well as the ordinary ones
there are some of my stocks in the liq N2 which came stably transfected
with an EBNA gene and which I suppose might behave oddly although
off-hand I can't think of a reason why they should.
> Theodosios Kyriakou


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