ways to get around self activation in yeast 2 hybrid...
kirkpat at ?cc.umanitoba.ca?
Mon Jun 29 17:04:54 EST 1998
In article <3592BE36.B916605F at bs.jhmi.edu>, Sai Iyer <siyer at bs.jhmi.edu> wrote:
> hail all,
> my construct is a strong self activator when i use it as a BD
> 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
During a review of two-hybrid articles, I came upon a promising one in
which they "dampened" the activation, then compared the resulting "true"
activation to the dampened state. If none of the other methods suggested
to date work for you and you really want to use the clone you have, you
may want to check out this paper:
Cormack and Somssich. Analytical Biochemistry. 248: 184-186 (1997)
I've never tried this out and I don't know anyone personally who has - it
could be more trouble than it's worth or it may be your life-saver...
Happy protein hunting!
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