Sensitive and Convenient Yeast Reporter Assay for High-Throughput Analysis by Using a Secretory Luciferase from Cypridina noctiluca

摘要

The yeast reporter assay has been widely used innvarious applications such as detection of endocrinendisruptors and analysis of protein-protein interactionsnby the yeast two-hybrid system. The molecular characteristicsnof the reporter enzyme are critical determinantsnfor this assay. We herein report the establishment of annovel yeast reporter assay using a secretory luciferase,nCypridina noctiluca luciferase (CLuc), as an alternativento the conventional u0001-galactosidase. The CLuc reporternassay in yeast is more sensitive and convenient thannthe conventional assay. A yeast high-throughput reporternassay was established with a laboratory automationnsystem, and the transcriptional activity of hundredsnof yeast promoter fragments was comprehensively determined.nOur results indicate that the yeast CLucnreporter assay is a promising tool for large-scale andnsensitive analysis in the development of new drugs andnin various fields of biotechnology research.