Two Hybrid False Positives?

Erica Golemis EA_Golemis at fccc.edu
Thu Nov 21 12:38:25 EST 1996


If you can stand some handwaving:
The vanishing blue phenotype is not that uncommon.  We've been
characterizing various behaviours of supposed false positives, and one
thing we've found is that a number of them are related to a peculiarity
of protein expression.  That is, if you take the initial false positive,
and retransform it with the protein you selected it with, a high
percentage of the yeast colonies arising will not express either the
bait protein, or the interactor protein, even though both plasmids are
present.  By high percentage, I'm talking ~ 80-90%.  Our current guess
is that these interactors, either alone or in combination with a
particular bait, is doing something that the yeast do not like, so they
are getting rid of it.  On the other hand, in the original isolation of
these interactors, you had previously applied a very strong selection in
favor of expression of both halves of the combination, ie the ability to
grow in the face of an amino acid deprivation - which incidentally fits
with Joe's phenomenon. 
Erica



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