Interaction mating methods in two-hybrid systems

Yeast two hybrid interaction mating methods. PDF version of the reprint from Finley lab at Wayne State University.

Reference:

Methods in Enzymology, Applications of Chimeric Genes and Hybrid Proteins, Part C, Protein-Protein Interactions and Genomics, eds John Abelson, Scott Emr, and Jeremy Thorner, vol 328, pp26-46, 2000

Content

Introduction

Isolating new interactors from libraries

Constructing and characterizing the bait strain

Preparing the pretransformed library and control strains

Screening for interactors by mating

Characterizing interactors

Testing interactions between small sets of proteins by cross-mating assay

Mapping networks of interacting proteins

Elaborating protein networks by reiterative interactor hunts

Constructing new bait strains by recombination cloning

Screening two-hybrid arrays

Exploring the functions of individual protein interactions

Isolating interaction mutants and suppressors

Breaking interactions with proteins and peptides

Isolating disruptive peptide aptamers

Interpreting results

Concluding remarks

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