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

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journal contribution
posted on 01.07.2010 by Yuki Tochigi, Natsuko Sato, Takehiko Sahara, Chun Wu, Shinya Saito, Tsutomu Irie, Wataru Fujibuchi, Takako Goda, Ryoichi Yamaji, Masahiro Ogawa, Yoshihiro Ohmiya, Satoru Ohgiya
The yeast reporter assay has been widely used in various applications such as detection of endocrine disruptors and analysis of protein−protein interactions by the yeast two-hybrid system. The molecular characteristics of the reporter enzyme are critical determinants for this assay. We herein report the establishment of a novel yeast reporter assay using a secretory luciferase, Cypridina noctiluca luciferase (CLuc), as an alternative to the conventional β-galactosidase. The CLuc reporter assay in yeast is more sensitive and convenient than the conventional assay. A yeast high-throughput reporter assay was established with a laboratory automation system, and the transcriptional activity of hundreds of yeast promoter fragments was comprehensively determined. Our results indicate that the yeast CLuc reporter assay is a promising tool for large-scale and sensitive analysis in the development of new drugs and in various fields of biotechnology research.

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