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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

Gateway-compatible yeast one-hybrid screens.

Bart Deplancke1, Vanessa Vermeirssen, H Efsun Arda

  • 1Program in Gene Function and Expression and Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605, USA.

CSH Protocols
|April 10, 2012
PubMed
Summary

The yeast one-hybrid system identifies protein-DNA interactions (PDIs) by screening DNA baits against protein preys. This high-throughput method detects interactions crucial for understanding transcription regulatory networks.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Protein-DNA interactions (PDIs) are fundamental to gene regulation.
  • Transcription factors (TFs) bind to specific DNA sequences to control gene expression.
  • Identifying these interactions is key to understanding regulatory networks.

Purpose of the Study:

  • To describe the Gateway-compatible yeast one-hybrid (Y1H) system.
  • To highlight its utility as a high-throughput, gene-centered method for PDI discovery.
  • To explain the mechanism for identifying interactions between DNA elements and proteins.

Main Methods:

  • Utilizes a yeast one-hybrid (Y1H) system with Gateway cloning.
  • DNA baits (cis-regulatory elements/promoters) are fused to HIS3 and lacZ reporter genes.
  • Yeast strains are screened for reporter gene expression upon prey protein binding to DNA bait.

Main Results:

  • The Y1H system allows for high-throughput screening of PDIs.
  • It can identify interactions involving both activators and repressors.
  • The method is gene-centered, starting with DNA baits.

Conclusions:

  • The Gateway-compatible Y1H system is an effective tool for discovering PDIs.
  • This method facilitates the study of transcription regulatory networks.
  • It enables the identification of novel TF-DNA binding events.