Best cell line for expression cloning but not COS-7 c

Ian A. York iayork at panix.com
Tue Jan 6 11:31:11 EST 1998


In article <grund-0601981652490001 at 129.194.96.102>,
Christophe Grundschober <grund at cmu.unige.ch> wrote:
>
>I tested COS-7 cells which are easely transfected but unfortunately they
>already express the gene I'm looking for. I'm now looking for other cell
>lines which could be used. They have to be easely transfected (by
>electroporation or other) and grow well. Does anybody have a suggestion?

If you're planning on doing transient expression for your assays, you
don't have a huge number of options.  You might be able to use WOP cells,
which are mouse cells transformed with the polyoma virus large T antigen; 
they're thus analogous to COS cells, and will amplify plasmids which
contain the polyoma origin of replication.  Depending on your library, you
might already have the polyoma ori; plasmids derived from pCDM8, which is
popular for library cloning, have both the SV40 origin and the polyoma
origin.   I've done a couple of transfections into WOP and got decent
levels of expression, though not as high as in COS.  

If you can use plasmids with EBNA1 and EBV origin of replication, such as
Invitrogen's pREP* series, you can get stable maintenance of plasmids in
primate or canine cell lines.  I don't know how easily transfectable
canine cell lines are, but you may be able to use something like 293 or
HeLa cells, which are quite transfectable. 

Good luck.

-- 
    Ian York   (iayork at panix.com)  <http://www.panix.com/~iayork/>
    "-but as he was a York, I am rather inclined to suppose him a
     very respectable Man." -Jane Austen, The History of England



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