The Yeast Two-Hybrid System: A Tool for Mapping Protein–Protein Interactions

Abstract

Virtually all processes in living cells are dependent on protein–protein interactions (PPIs). Understanding PPI networks is thus essential for molecular biology and disease research. One powerful genetic system for mapping PPIs both at a small scale and in a high-throughput manner is the yeast two-hybrid (Y2H) screen. In Y2H screening, PPIs are detected through the activation of reporter genes responding to a reconstituted transcription factor. In this introduction, we describe library- and array-based Y2H methods and explain their basic theory. We also include the rationale behind different Y2H approaches and strategies for optimizing results.