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Dear colleagues,
I'm a new graduate student working on yeast and have some problems with my project. I'm trying to identify yeast factors critical for a two-hybrid interaction. We are studying an interaction between two proteins, and we believe the interaction is dependent on phosphorylation of one of the partners. We'd like to identify the kinase responsible. I plan to use the yeast deletion library and identify those strains in which the two-hybrid interaction is interrupted. However, my experiments require three plasmids (the bait, the target and the reporter gene). Because there are more than 6,000 strains in the library, I don't want to transform all three plasmids into each strain. Is there anybody who is also using this library and/or screening it by the two-hybrid method? I'd like to have your suggetions as to how I can reduce the work. Any suggestions will be very much appreciated. Thanks!
Best wishes
Junbiao Dai
MCDB program
Iowa State University
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