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.

R

-- 
Richard P. Grant MA DPhil          
Structural Studies Group, MRC-LMB
http://www2.mrc-lmb.cam.ac.uk/personal/rpg/index.html
Please reply to rpg 'at' mrc-lmb.cam.ac.uk




More information about the Proteins mailing list