background in 2 hybrid screen

Ingrid Wolf iwolf at fhcrc.org
Sun Oct 22 10:38:17 EST 1995


I have cloned a plasmid encoding a LexA-fusion protein which I want to 
use for the yeast two hybrid screen. This bait-plasmid alone or together 
with the empty VP16 vector allows already growth of the transformed 
yeast clones on plates lacking His but they stay white on X-gal 
plates.With a positiv control, a known interacting protein fused to 
VP16,  the yeast transformants grow on plates lacking His and turn to 
dark blue on X-gal plates. The background growth of the bait on plates 
lacking His can not be suppressed by 3-aminotriazole (I titrated all 
concentrations up to 200 mM). My question is:
 Has anybody made a two hybrid screen by monitoring the interaction only 
with the blue color?
Please give me a message to:      iwolf at fhcrc.org
Thank you.





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