Yeast Protein–Protein Interaction Assays and Screens

S. de Folter, G.H. Immink

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

43 Citations (Scopus)


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.
Original languageEnglish
Title of host publicationPlant Transcription Factors, Methods in Molecular Biology
EditorsL. Yuan, S.E. Perry
Number of pages21
Publication statusPublished - 2011

Publication series

NameMethods in Molecular Biology
PublisherSpringer Science


  • complexes
  • GAL4 system
  • Protein-protein interactions
  • transcription factors
  • Y2H
  • yeast two-hybrid
  • YnH


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