High-throughput yeast two-hybrid assays for large-scale protein interaction mapping

Methods. 2001 Jul;24(3):297-306. doi: 10.1006/meth.2001.1190.


Protein-protein interactions play fundamental roles in many biological processes. Hence, protein interaction mapping is becoming a well-established functional genomics approach to generate functional annotations for predicted proteins that so far have remained uncharacterized. The yeast two-hybrid system is currently one of the most standardized protein interaction mapping techniques. Here, we describe the protocols for a semiautomated, high-throughput, Gal4-based yeast two-hybrid system.

MeSH terms

  • Culture Media
  • DNA, Recombinant / metabolism
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Gene Library
  • Genes, Reporter / genetics
  • In Vitro Techniques
  • Polymerase Chain Reaction
  • Proteins / chemistry*
  • Saccharomyces cerevisiae / genetics
  • Transformation, Genetic
  • Two-Hybrid System Techniques*


  • Culture Media
  • DNA, Recombinant
  • Fungal Proteins
  • Proteins