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

UMMS Affiliation

Program in Gene Function and Expression; Program in Molecular Medicine



Document Type


Medical Subject Headings

Culture Media; DNA, Recombinant; Fungal Proteins; Gene Library; Genes, Reporter; Polymerase Chain Reaction; Proteins; Saccharomyces cerevisiae; Transformation, Genetic; *Two-Hybrid System Techniques


Genetics and Genomics


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.

Rights and Permissions

Citation: Methods. 2001 Jul;24(3):297-306. Link to article on publisher's site


At the time of publication, Albertha J. Marian Walhout was not yet affiliated with the University of Massachusetts Medical School.

Related Resources

Link to Article in PubMed