In article <199807031633.MAA12036 at mail3.uts.ohio-state.edu>,
Ahnert.1 at osu.edu (Peter Ahnert) wrote:
> Thank you very much for your reply. This is pointing me in the right
> direction. Basically we are looking for any sort of vector that will allow
> expression of a foreign protein in a mammalian cell to see what happens
> in terms of posttranslational modifications. The problem is that we sort
> of need a place to start and could use any promoter that will give good
> expression.
> Thanks again for your suggestions.
> Peter Ahnert.
Glad my earlier post was of some help. My personal favourite
is pCI-neo from Promega but I've also had reasonable results
with Stratagene's pBK-CMV and colleagues give Invitrogen's
pcDNA series the thumbs up. I don't know if you are limited
as far as your choice of mammalian cells is concerned, but a
CMV-driven vector in COS cells is an excellent combination
because of the viral upregulation of the promoter. COS
cells are also nice and easy to transfect (eg. calcium phosphate).
Since you are interested in post-translational modification you
may also want to consider in vitro transcription/translation
(eg. Promega's TnT kit) as this may be a more direct approach.
Good luck,
Bernard
[No commercial affiliation - and if Promega are watching this,
I hope you'll eventually get around to supplying the twice-promised
TnT information]
--
Bernard Murray, PhD
Dept. Cell. Mol. Pharmacol., UCSF, San Francisco, USA
