ways to get around self activation in yeast 2 hybrid...

Bernard Murray bpmurray*STUFFER* at socrates.ucsf.edu
Tue Jun 30 21:12:26 EST 1998


In article <359782E8.7F617793 at wadsworth.org>, Brian Cohen
<cohen at wadsworth.org> wrote:

> There's actually an interesting system from Stephen Elledge's and
Michael Karin's
> Labs that uses a membrane based interaction trap- no need to worry about self
> activating proteins, since the whole system is outside of the nucleus. 
The only
> down side is that you have to move your bait to a new vector, and, at
least last
> time I corresponded with them about the system, there were no libraries
available.
> However, that was six months ago, so there may have been some changes
since then.
> The reference is:
>     Aronheim et. al. Mol Cell Bio 17:3094-3102
> I haven't used teh system myself, but I wonder if anyone here has used
the system
> and could comment on it?
> Hope this helps,
> Brian

I believe that Stratagene are just about to start selling this.
I've only seen a brief presentation by one of their reps but
it looked intriguing.  Yes, you have to subclone your
sequences into a vector that allows fusion with a myristoylation
sequence.  Looks to have good potential for large proteins,
transcription factors and possibly for membrane-bound proteins.
Sorry I can't give any practical input and I also look forward
to seeing other peoples' results.
          Bernard
-- 
Bernard Murray, PhD
Dept. Cell. Mol. Pharmacol., UCSF, San Francisco, USA



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