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Updated: Jul 23, 2025

A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions
Published on: June 6, 2018
Next-Generation Yeast Two-Hybrid Screening to Discover Protein-Protein Interactions
J Mitch Elmore1,2,3, Valeria Velásquez-Zapata4,5, Roger P Wise6,4,5
1USDA-Agricultural Research Service, Cereal Disease Laboratory, St. Paul, MN, USA. mitch.elmore@usda.gov.
Abstract:
Yeast two-hybrid is a powerful approach to discover new protein-protein interactions. Traditional methods involve screening a target protein against a cDNA expression library and assaying individual positive colonies to identify interacting partners. Here we describe a simple approach to perform yeast two-hybrid screens of a cDNA expression library in batch liquid culture. Positive yeast cell populations are enriched under selection and then harvested en masse. Prey cDNAs are amplified and used as input for next-generation sequencing libraries for identification, quantification, and ranking.
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