Yeast Two-Hybrid System and Membrane Proteins

Doug Johnson djohnson at science.uottawa.ca
Sat Nov 22 09:57:41 EST 1997


Colleagues:

I am  potential  user of the two-hybrid system. In the literature a 
wide variety of interactions have been studied but I could only find 
one reference to an interaction involving a membrane bound protein. 
(Zha et al. 1996, J. Biol. Chem. 271: 7440-7444). I am sure there 
are many others. The authors removed the transmembrane(TM) domains in 
order to avoid "problems with targeting of proteins to the nucleus". 

The question that I have for your comment: " is removal necessary?". 
Given that the cDNA clones may not be full-length, perhaps for an 
efficient cloning there will be some clones that lack the  TM domain 
if it is amino-terminal. Conversely, it the TM domain is 
carboxy-terminal, would a random primed library suffice?

Any help cheerfully received.

Doug Johnson
djohnson at science.uottawa.ca



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