Title

A high-throughput system for two-hybrid screening based on growth curve analysis in microtiter plates

Document Type

Article

Publication Date

2003

Publication Title

Analytical Biochemistry

Volume

316

Issue

2

First Page

171

Last Page

174

DOI

10.1016/S0003-2697(02)00706-6

Abstract

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.

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