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GAL4 nuclear localization signal

George Theodoris ez003450 at bullwinkle.ucdavis.edu
Mon Oct 24 11:46:13 EST 1994

Colin Rasmussen (ras-1 at bones.biochem.ualberta.ca) wrote:

: My question is why would you want to do this. If it's to purify a protein
: there are lots of other ways. The two-hybrid idea depends on nuclear
: localization.
I want to know if my protein is functional when fused to GAL4. Since my 
protein is cytoplasmic, I can't verify that the fusion is functional as 
long as it is targeted to the nucleus. My worry is that when you fuse a 
protein to GAL4 you don't know what effect that will have on the 
conformation of the protein. Perhaps fusing it to GAL4 will disrupt the 
confirmation, rendering it unable to interact with whatever proteins it 
interacts with. If my protein is not functional when fused to GAL4 I'll 
know that it may not be able to interact with the proteins it normally 
interacts with and that the two-hybrid system probably isn't the best
way of identifying interacting proteins. -George

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