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two-hybrid cloning

Andre Nantel nantel at bri.nrc.ca
Fri Jun 28 09:25:23 EST 1996

In article <31D339E4.6333 at cfi.unsw.edu.au>
Rick Nicholson <R.Nicholson at cfi.unsw.edu.au> writes:

> Hi,
>         Does anyone know if yeast based two-hybrid cloning systems such as 
> Clontech's Matchmaker system have ever been successful in cloning 
> receptors by using ligand as the target?
> Please let me know if anyone has tried and been successful, or even if 
> anyone has tried unsuccessfully.
> Thanks, Rick.

Enzyme-substrate interactions have occasionnaly been reported but you
usually have to mutate the ligand/bait in such a way as to prevent the
enzymatic reaction from being completed. For example, you can check the
paper by Wu et al. (JBC, 271:3265, 1996) => Interaction between the
kinases RAF and its substrate MEK are only observed when the serine
phosphorylation sites on MEK are mutated to alanines.

Andre Nantel                             Purveyor of fine genetically-
Biotech Research Institute               engineered roadkills for the
National Research Council Canada         Information Superhighway
Montreal, Quebec
nantel at biotech.lan.nrc.ca or andre at bri.nrc.ca

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