An introduction to Yeast Two-Hybrid

The two-hybrid assay is a well-established system to analyze protein-protein interactions. We offer to provide a pooled version of the set of yeast transformants containing open reading frames (ORFs) fused to the Gal4 activation domain derived from individual PCRs and transformation of the entire set of yeast ORFs. This pool was generated by combining the array of ~6000 strains as described in Uetz et al. 2000. We will provide as well the relevant vectors and yeast strains. Any protein of interest can be constructed as a DNA-binding domain hybrid in a compatible vector and used to screen this pool to identify potential interacting partners; positives can be selected by their ability to grow in the absence of histidine and/or adenine.