Membrane proteins in dihybrid?

SLForsburg susan_forsburg at qm.salk.edu
Sat Dec 10 12:12:28 EST 1994


"Terri G. Kinzy" <tk2h+ at andrew.cmu.edu> wrote:
>
> (snip)
>
> Particularly, this person wants to find the receptor for his protein. 
> Has anyone tried this or looked specifically at pairs of proteins, one
> of which is presumed or known to be transmembrane.  I guess the
> questions are (1) transport into the nucleus, would these types of
> protein get stuck or (2) would they not be properly translated in the
> cytoplasm or unable to fold, etc..

I would expect that in cases of this sort, truncations are the way to
go.  Presumably a cDNA including the interaction domain(s) would be
sufficient to allow interaction in the two hybrid system;  this would 
require a random-primed cDNA library in the prey plasmid.  Yet 
another example of the basic concept that two-hybrid screening
is a tool, and not necessarily reflecting normal protein function.
After all, if you sent your fusions to the membrane, you'd never 
get activation...

susan
susan_forsburg at qm.salk.edu



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