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.
Diaz-Camino
Claudia
C
Gene Expression Group, Plant Biotechnology Institute, National Research Council Canada, 110 Gymnasium Place, Saskatoon, SK, Canada S7N 0W9.
Risseeuw
Eddy P
EP
Liu
Enwu
E
Crosby
William L
WL
D000818:Animals; D029681:Arabidopsis Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D001331:Automation; D002999:Clone Cells; D003124:Colorimetry; D010957:Plasmids; D011485:Protein Binding; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics / Q000254:growth & development*
D029681:Arabidopsis Proteins