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One-hybrid cloning

KOS at NL.UVA.AMC KOS at NL.UVA.AMC
Mon Oct 11 09:34:37 EST 1993


Dear yeastnetters,
 
I am trying to clone a YEAST transcription factor with a
procedure based on the two-hybrid system. It actually is a one-
hybrid system because I only use the library with the GAL4
activation domain to find a factor that is binding not to a GAL4
site but to my binding site of interest. I have experienced some
dificulties with this procedure. Therefore my questions are: has
someone tried this, was the procedure successful, what were the
problems and how were they solved? Note that I am looking for
factors that are endogenous in yeast! The only successfull
reports I have heard sofar were with factors from higher
eucaryotes, without interference of an endogenous background.
Please send your answers directly to me at:
KOS at AMC.UVA.NL
I will later summarize the answers for the newsgroup.
Thank you for your attention,
 
Arnoud Kal
Dept. for Biochemistry
University of Amsterdam
The Netherlands



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