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Phosphorylation of target proteins in yeast two-hybrids

Andre Nantel nantel at bri.nrc.ca
Wed Jul 3 13:27:17 EST 1996


In article <4rd6rp$54l at seagoon.newcastle.edu.au>
<@medicine-dmb.newcastle.edu.au> writes:

> Howdy,
> 
> I am using a yeast two-hybrid system to screen for proteins that 
> interact with my target protein. I have used a full-length target 
> protein with no success. I want to try to phosphorylate this protein in 
> the yeast cell to see if this modification will induce protein 
> interactions.
> 
> If anyone can help me with suggestions as how to go about this, I'd be 
> tickled. I am looking for things like when, how, what and how much.
> 
> Thanks,
> 
> 
> Glenn

You may consider mutating the putative phosphorylation sites in your
target protein (JBC, 271:3265) or using the Tribrid system
(Biotechnology, 13:1474).

Have fun.

----------------------------------------------------------------------
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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