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Related Concept Videos

Genetic Screens02:46

Genetic Screens

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Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which...
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Related Experiment Video

Updated: Mar 10, 2026

A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions
14:23

A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions

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Performing Yeast One-Hybrid Library Screens.

Juan I Fuxman Bass1, John S Reece-Hoyes1, Albertha J M Walhout1

  • 1Program in Systems Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01605.

Cold Spring Harbor Protocols
|December 10, 2016
PubMed
Summary

The yeast one-hybrid (Y1H) assay identifies transcription factor (TF) DNA-binding interactions. This method screens TF libraries against a DNA bait to find specific TF-DNA binding events.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Yeast one-hybrid (Y1H) assays are a standard method for identifying transcription factor (TF) DNA-binding interactions.
  • These assays utilize a yeast strain engineered with a reporter gene linked to a specific DNA bait sequence.

Purpose of the Study:

  • To describe a detailed protocol for performing a Y1H screen to identify novel TF-DNA interactions.
  • To provide a method for screening cDNA libraries against a DNA bait sequence.

Main Methods:

  • The Y1H assay involves transforming a yeast bait strain with a library of cDNA fragments cloned into a plasmid expressing each cDNA as a fusion protein with the Gal4 activation domain (AD).
  • Reporter gene activation (HIS3 and LacZ) indicates a TF-DNA interaction.
  • Positive interactions are retested and the interacting TF is identified by sequencing the cDNA insert.

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Mating-based Overexpression Library Screening in Yeast
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Mating-based Overexpression Library Screening in Yeast

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Enhanced Yeast One-hybrid Screens To Identify Transcription Factor Binding To Human DNA Sequences
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Enhanced Yeast One-hybrid Screens To Identify Transcription Factor Binding To Human DNA Sequences

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Related Experiment Videos

Last Updated: Mar 10, 2026

A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions
14:23

A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions

Published on: June 6, 2018

14.3K
Mating-based Overexpression Library Screening in Yeast
11:39

Mating-based Overexpression Library Screening in Yeast

Published on: July 6, 2018

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Enhanced Yeast One-hybrid Screens To Identify Transcription Factor Binding To Human DNA Sequences
11:25

Enhanced Yeast One-hybrid Screens To Identify Transcription Factor Binding To Human DNA Sequences

Published on: February 11, 2019

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Main Results:

  • The described Y1H screening method successfully identifies TF-DNA binding partners.
  • The protocol allows for the screening of both commercial and in-house generated cDNA libraries.

Conclusions:

  • The yeast one-hybrid assay provides a robust platform for discovering transcription factor-DNA binding specificities.
  • This protocol facilitates the identification of TFs that interact with specific DNA regulatory elements.