Yeast Transformation/Two-Hybrid System
trotta at seqvax.caltech.edu
Tue Sep 28 17:20:00 EST 1993
I am currently trying to get a high efficiency transformation of yeast
with two plasmids. I have only been able to get as high as 1x10^3
transformants/ug DNA. This is not high enough to screen a library.
The second part of my post is to get general ideas on the two-hybrid
system. Stuff like which strains have worked best, methods of
screening, what hasn't worked etc. I have begun screening the
libraries in a His reporter strain YPB2 and have not had any success.
I also used a LexA derived system without any success either. It
seems as though there are quite a few false positives in each. In general
I have recovered 40-50 positive clones which by themselves can activate
transcription. I am interested in hearing about other screens.
trotta at caltech.cco.edu
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