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 beta-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 beta-galactosidase assay. We conclude that the method presented here is reproducible, of equal or greater sensitivity than the beta-galactosidase assay, and can be further adapted for application to the conduct of large-scale, automated yeast two-hybrid experiments.
Mesh Terms:
Animals, Arabidopsis Proteins, Automation, Clone Cells, Colorimetry, Plasmids, Protein Binding, Reproducibility of Results, Saccharomyces cerevisiae, Two-Hybrid System Techniques, Yeasts
Anal. Biochem.
Date: May. 15, 2003
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