Examining the function of proteins with the yeast two-hybrid system

Review and detailed protocol written by Dr. Russ Finley at Finley lab, Wayne State University School of Medicine.

The yeast two-hybrid system provides a relatively straight forward approach to understanding protein function. Section II outlines the basic components of the interaction trap, a yeast two-hybrid system developed in the Brent lab (Gyuris et al., 1993). More detailed background information can be obtained in a number of recent reviews (Ausubel et al., 1987-1996; Finley and Brent, 1995; Mendelsohn and Brent, 1994). Section III contains an interactor hunt protocol, which is a condensed and updated version of the original protocol we first posted on the Internet in 1992 and subsequently updated (Finley and Brent, 1995; Finley et al., 1997). The version presented here is the one we currently use in our lab and represents our attempts to streamline and scale up the these techniques to facilitate characterization of large networks of interacting proteins. It is also useful for individual hunts. Section IV discusses alternative approaches specifically designed to look at large protein networks; the ultimate goal of developing these and related approaches is to eventually map all of the interactions encoded by a genome. Section V discusses briefly two-hybrid approaches to understanding the functions of individual protein interactions.

Last update 05-Mar-2005, Rating n/a of 0 votes.