ABSTRACT
The yeast two��hybrid system is a procedure for the detection of protein protein interactions in recombinant Saccharomyces cerevisiae. The system has been described in detaii others (Fields and Song, 1989, 1 Chien et aI. , 99 1 ; Fritz and Green, 1992; Bartel et al. , 1993 a and b; Fields and Sternglanz, 1 994; Phizicky and Fields , 1 995). This chapter is intended to provide a practical account for the application of the two-hybrid system in a typical molecular biology laboratory set ting. We have tried to emphasize experimental conditions which, in our experience, arc helpful for minimizing the recovery of artefacts with this system. While our own practical experience has concentrated on the identification of protein-interaction partners in two-hybrid cDNA expression libraries of A rabidopsis thaliana, the pro tocols should prove generally useful for the construction and screening of libraries from other organisms.
