Yeast two-hybrid screen

Methods Enzymol. 2014:539:31-51. doi: 10.1016/B978-0-12-420120-0.00003-7.

Abstract

Yeast two-hybrid is a method for screening large numbers of gene products (encoded by cDNA libraries) for their ability to interact with a protein of interest. This system can also be used for characterizing and manipulating candidate protein: protein interactions. Interactions between proteins are monitored by the growth of yeast plated on selective media.

Keywords: Activation domain (AD) fusion protein; Back-transformation of yeast; Cloning and transformation methods; Confirmation of positive interactors; DNA-binding domain (DB) fusion protein; Electroporation of E. coli; Plasmid rescue; Self-activation of pDBLeu-X (and pPC86-Y); Small-scale yeast transformation; Yeast two-hybrid screen; pDBLeu-X.

MeSH terms

  • Culture Media
  • Escherichia coli
  • Protein Binding
  • Saccharomyces cerevisiae / growth & development*
  • Two-Hybrid System Techniques*

Substances

  • Culture Media