GST/His Tag expression woes
Richard P. Grant
rgrant at netscape.net
Thu Mar 9 09:00:26 EST 2000
In article <8a670b$sb8$1 at phobos.scripps.edu>, "Antonin Tutter"
<atutter at aim.salk.edu> wrote:
> > I have not varied the length of induction, nor the concentration of
> > IPTG. Can anyone
> > out there offer any suggestions as to which conditions to vary. Is this
> > doomed to fail?
> > Should I start looking at baculovirus or gateway technology?
> you just suggested the right things to do. it sounds like the fusion is
> toxic to the cells at the current induction conditions,
I can't see the original poster's post, but with a toxic construct it's
a good idea to use a strongly repressing strain (e.g. pLysS) so that you
get good growth (i.e. you're preventing the bad effects from a leaky
repressor), and then induce as normal.
Richard P. Grant MA DPhil
Structural Studies Group, MRC-LMB
Please reply to rpg 'at' mrc-lmb.cam.ac.uk
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