Yeast two-hybrid approaches to dissecting the plant defense response

Methods Mol Biol. 2007:354:79-83. doi: 10.1385/1-59259-966-4:79.

Abstract

We describe a reliable GAL4-based yeast two-hybrid system for identifying and isolating clones encoding proteins interacting with a protein of interest. This two-hybrid system gives extremely low background and few false-positive clones, making it ideal for library screening purposes. We have successfully used it not only to isolate Arabidopsis NPR1-interactors from rice but also to pull out the rice NPR1 ortholog using one of the interactors as bait.

MeSH terms

  • Arabidopsis / genetics
  • Arabidopsis / immunology
  • Arabidopsis / metabolism*
  • DNA, Plant / isolation & purification
  • Gene Library
  • Immunity, Innate / physiology*
  • Oryza / genetics
  • Oryza / immunology
  • Oryza / metabolism*
  • Plant Diseases*
  • Plasmids / isolation & purification
  • Transformation, Genetic
  • Two-Hybrid System Techniques*
  • Yeasts / cytology
  • beta-Galactosidase / metabolism

Substances

  • DNA, Plant
  • beta-Galactosidase