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

Brian Cohen cohen at wadsworth.org
Mon Jun 29 07:04:56 EST 1998


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

Sai Iyer wrote:

> hail all,
>
>         my construct is a strong self activator when i use it as a BD fusion to
> fish interacting proteins from a cDNA library.  use of 3 amino triazole
> is not a option since the activation is immediate (in context to 2
> hybrid activation times) i.e in about 5 min.  so is there any other
> possible option that i can employ to fish out interacting proteins from
> a library using 2 hybrid?? i should mention that i dont have
> any candidate clones yet; so switching BDs and ADs is also not an option
> (yet)...thanks
>
> --
> Sai Iyer (siyer at bs.jhmi.edu)
> Pre-doctoral fellow i.e lowly graduate student
> Dept. of Biological Chemistry
> Johns Hopkins University School of Medicine



--
Brian D. Cohen, Ph.D.
Laboratory of Reproductive and Metabolic Disorders
David Axelrod Institute #5031
Wadsworth Center
NYS Dept. of Health
P.O. Box 22002
Albany, NY 12008-2002
(518) 474-4172
cohen at wadsworth.org

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