UNIT 12.12 Yeast One-Hybrid Screening for DNA-Protein Interactions
Published Online: 1 AUG 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Ouwerkerk, P. B. and Meijer, A. H. 2001. Yeast One-Hybrid Screening for DNA-Protein Interactions. Current Protocols in Molecular Biology. 55:12.12.1–12.12.12.
- Published Online: 1 AUG 2001
- Published Print: JUL 2001
One-hybrid screening in yeast is a powerful method to rapidly identify heterologous transcription factors that can interact with a specific regulatory DNA sequence of interest (the bait sequence). In this technique, the interaction between two proteins (bait and prey) is detected via in vivo reconstitution of a transcriptional activator that turns on expression of a reporter gene. Detection is based on the interaction of a transcription factor (prey) with a bait DNA sequence upstream of a reporter gene. To ensure that DNA binding results in reporter-gene activation, cDNA expression libraries are used to produce hybrids between the prey and a strong trans-activating domain. The advantage of cloning transcription factors or other DNA-binding proteins via one-hybrid screenings, compared to biochemical techniques, is that the procedure does not require specific optimization of in vitro conditions.