Investigation of Light-Regulated Protein–Protein Interactions Using Yeast Two-Hybrid Assays

Abstract

Yeast two-hybrid allows for investigation of interactions between two proteins in vivo. Yeast media can be supplemented with phycocyanobilin, allowing the formation of photoactive phytochromes, and the investigation of their light-regulated interactions. Three reporter assays are described; the histidine growth selection assay for the HIS3 reporter, and the filter lift and quantitative ONPG assays for the LacZ reporter. Design considerations for yeast two- and three-hybrid experiments are also covered.