pCMV-Neo resistance

[John Dixon]

In article <1996Jun6.192612.26489 at alw.nih.gov>, bernard at elsie.nci.nih.gov
(Bernard Murray) wrote:

> In article <31B6B93A.7AD5 at urz.uni-heidelberg.de>, 
> rico.laage at urz.uni-heidelberg.de says...
> >
> >Graham Atherton wrote:
> >> 
> >> Does anyone know the whereabouts, commercial or otherwise, of a plasmid
> >> containing G418 - resistance gene driven by the CMV promoter.
> >> I have searched all of the catalogues that come to mind with no success!

snip

>         The neo gene seems pretty efficient...

snip

Watch out for old (pre-1990) neo cassettes as they may carry a mutation
which converts a G to a T resulting in an glu to asp alteration at codon
182. The mutation abolishes an XhoII rgatcy-not xhoI- site and is
therefore easy to check for. See ref for more info: Yenofsky et al, PNAS
1990, 87, 3435-3439, a mutant neo gene reduces the resistance of
transformants to antibiotic selection pressure.

I have found that the corrected version gives me approx 7 times as many
G418R murine ES cells when compared with the mutant (unpublished
observation).

Cheers


John

-- 
John Dixon                                        Lab 44 (1223) 334131
Wellcome/CRC Institute                            Fax 44 (1223) 334134
Department of Genetics
Cambridge University    
United Kingdom                           e-m: jpcd0 at mole.bio.cam.ac.uk