Plant Transcription Factors pp 145-165

Part of the Methods in Molecular Biology book series (MIMB, volume 754)

Yeast Protein–Protein Interaction Assays and Screens

Protocol

Abstract

Most transcription factors fulfill their role in protein complexes. As a consequence, information about their interaction capacity sheds light on a protein’s function and the molecular mechanism underlying this activity. The yeast two-hybrid GAL4 (Y2H) assay is a powerful method to unravel and identify the composition of protein complexes. This in vivo based system makes use of two functional protein domains of the GAL4 transcription factor, each fused to a protein of interest. Upon interaction between the two proteins under study, a transcriptional activator gets reconstituted and reporter genes get activated, allowing the yeast to grow on selective medium. In this chapter protocols are given for Y2H library screening, directed Y2H screening, Y2H matrix screening, and YnH screening involving more than two proteins.

Key words

Protein–protein interactions yeast two-hybrid Y2H YnGAL4 system complexes transcription factors 

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Laboratorio Nacional de Genómica para la Biodiversidad (LANGEBIO)Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN)Irapuato, GuanajuatoMéxico
  2. 2.Plant Research International, Centre for BioSystems Genomics (CBSG)WageningenThe Netherlands

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