Methods protocols: Gene-Centered Yeast One-Hybrid Assays

December 16, 2016

Four protocols on methods and considerations when performing yeast one-hybrid assays are featured on Cold Spring Harbor Protocols.


An important question when studying gene regulation is which transcription factors (TFs) interact with which cis-regulatory elements, such as promoters and enhancers. Addressing this issue in complex multicellular organisms is challenging as several hundreds of TFs and thousands of regulatory elements must be considered in the context of different tissues and physiological conditions. Yeast one-hybrid (Y1H) assays provide a powerful "gene-centered" method to identify the TFs that can bind a DNA sequence of interest. In this introduction, we describe the basic principles of the Y1H assay and its advantages and disadvantages and briefly discuss how it is complementary to "TF-centered" methods that identify protein-DNA interactions for a known protein of interest.


Fuxman Bass JI, Reece-Hoyes JS, Walhout AJ. (2016). Gene-Centered Yeast One-Hybrid Assays. Cold Spring Harb. Prot., 2016(12).


Gene-Centered Yeast One-Hybrid Assays

Generating Bait Strains for Yeast One-Hybrid Assays


Performing Yeast One-Hybrid Library Screens


Zymolyase-Treatment and Polymerase Chain Reaction Amplification from Genomic and Plasmid Templates from Yeast


Colony Lift Colorimetric Assay for β-Galactosidase Activity


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Juan Fuxman Bass, PhD

Assistant Professor of Biology


Boston University

5 Cummington mall

Boston, MA 02215

e-mail: fuxman[at]

Phone: (617) 353-2448

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