The yeast two-hybrid system: a tool for mapping protein-protein interactions

Cold Spring Harb Protoc. 2015 May 1;2015(5):425-30. doi: 10.1101/pdb.top083345.

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.

MeSH terms

  • Protein Interaction Maps
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Two-Hybrid System Techniques*

Substances

  • Recombinant Proteins