Hi Nate,
There can be no consensus I guess, most importantly because transmembrane
proteins are so diverse :-)
We typically use transient transfection at the screening stage, and often for
large-scale expression (e.g. PMID 24909990). Bac-Mam has some advantages
(nicely described in 25299155, 27041595), but it's obviously more time
consuming. Lentiviral systems (we use Clontech) are also great.
But again the target often determines the choice of system. Whether it
contains multiple subunits, whether overexpression has a negative impact on
cell health (a tetO switch can be helpful, 12370422). Also, I'd use different
systems depending whether the recombinant protein is intended for functional,
fluorescence microscopy, structural analyses (say X-ray vs cryo-EM).....
Best wishes,
radu
--
Radu Aricescu
MRC Laboratory of Molecular Biology
Francis Crick Avenue
Cambridge Biomedical Campus
Cambridge CB2 0QH, U.K.
tel: +44-(0)1223-267049
fax: +44-(0)1223-268305
www: http://www2.mrc-lmb.cam.ac.uk/group-leaders/a-to-g/radu-aricescu
> Hello everyone,
>
> I wonder if there is a consensus for what is currently the best system to
> express transmembrane proteins in mammalian cells?
>
> I think that baculovirus transduction ("Bacmam" anf the likes) has been
> used historically more (?) but would like to know if the modern adenovirus
> systems offer any advantages in terms of expression levels. I used both in
> the past but not for transmembrane proteins and never compared them back to
> back for the same protein.
>
> Has anyone attempted a direct comparison? (It has to be mammalian cells, so
> baculovirus/insect cells won't do).
>
> Thanks for any comments/insights,
>
> Nate
>
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