Two hybrid Q.

[Erica Golemis]

Rafael Maldonado <rafael at corona> wrote:
>
> 
> 
> Hi folks!
> 
> I'm trying the two hybrid method for detecting interacting proteins in yeast.
> I have seen that the proteins to be tested are disposed in the C-terminal 
> of the gal4 protein domains (both). I mean, the fusions are always in the 
> N*gal4domain-yourfavoriteprotein*C direction.
> 
> Because I'm interested in the interaction with the N-terminal end of my 
> protein, can I use the fusion in the other way, N*Myprotein-gal4domain*C?
> 
> Have anyone tried that? Do you think it would work?
> 
> Thanks
> 
> Rafa
> 

       --Although not directly useful for people working with Clontech reagents,
there are available vectors to make fusions to LexA in which the LexA
domain would be located at the C-terminus of the fusion.  It can make a
remarkable difference in the properties of the fusion protein in some
cases where important residues are known to be located at the N-terminus
of the fusion domain.  Anyone wishing info can email me -
Erica