A high-throughput system for two-hybrid screening based on growth curve analysis in microtiter plates
The yeast two-hybrid system is a powerful tool for identifying novel protein-protein interactions. In general, biochemical marker genes such as lacZ are exploited for indirect quantification of the interaction, and commonly involve the conduct of rather laborious β-galactosidase assays. This paper describes a simple alternative method based on growth curve analysis of yeast cultures that is amenable to microtiter plate format, and therefore allows the quantification of large numbers of yeast two-hybrid combinations. The analyzed results of yeast cultures grown in microtiter plates were compared with those obtained from the classical β-galactosidase assay. We conclude that the method presented here is reproducible, of equal or greater sensitivity than the β-galactosidase assay, and can be further adapted for application to the conduct of large-scale, automated yeast two-hybrid experiments.
Diaz-Camino, Claudia; Risseeuw, Eddy P.; Liu, Enwu; and Crosby, William L., "A high-throughput system for two-hybrid screening based on growth curve analysis in microtiter plates" (2003). Analytical Biochemistry, 316, 2, 171-174.