Detecting Blue Light-Dependent Protein-Protein Interactions by LexA-Based Yeast Two-Hybrid Assay

Methods Mol Biol. 2021:2297:147-154. doi: 10.1007/978-1-0716-1370-2_15.

Abstract

The LexA-based yeast two-hybrid system is one of the most powerful techniques used to detect blue light-dependent protein-protein interactions. In Arabidopsis, many protein-protein interactions in blue light signaling pathway were identified using this system. Here we present an easy and efficient method of the LexA-based yeast two-hybrid assay for testing protein-protein interactions in a blue light-dependent manner.

Keywords: Blue light signaling pathway; Blue light-dependent; LexA; Protein–protein interaction; Yeast two-hybrid assay.

MeSH terms

  • Arabidopsis / metabolism
  • Arabidopsis Proteins / metabolism
  • Bacterial Proteins / metabolism
  • Color
  • Genetic Vectors
  • Leucine / metabolism
  • Light
  • Plasmids
  • Protein Interaction Mapping / methods*
  • Saccharomyces cerevisiae / genetics
  • Serine Endopeptidases / metabolism
  • Signal Transduction
  • Transformation, Genetic
  • Two-Hybrid System Techniques*
  • beta-Galactosidase / analysis

Substances

  • Arabidopsis Proteins
  • Bacterial Proteins
  • LexA protein, Bacteria
  • beta-Galactosidase
  • Serine Endopeptidases
  • Leucine