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

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• 作者：Yuki Tochigi ; Natsuko Sato ; Takehiko Sahara ; Chun Wu ; Shinya Saito ; Tsutomu Irie ; Wataru Fujibuchi ; Takako Goda ; Ryoichi Yamaji ; Masahiro Ogawa ; Yoshihiro Ohmiya ; Satoru Ohgiya
• 刊名：Analytical Chemistry
• 出版年：2010
• 出版时间：July 1, 2010
• 年：2010
• 卷：82
• 期：13
• 页码：5768-5776
• 全文大小：367K
• 年卷期：v.82,no.13(July 1, 2010)
• ISSN：1520-6882

文摘

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.