Sensitive and convenient yeast reporter assay for high-throughput analysis by using a secretory luciferase from Cypridina noctiluca.
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 beta-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.
Tochigi
Yuki
Y
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira-ku, Sapporo 062-8517, Japan.
Sato
Natsuko
N
Sahara
Takehiko
T
Wu
Chun
C
Saito
Shinya
S
Irie
Tsutomu
T
Fujibuchi
Wataru
W
Goda
Takako
T
Yamaji
Ryoichi
R
Ogawa
Masahiro
M
Ohmiya
Yoshihiro
Y
Ohgiya
Satoru
S
D000818:Animals; D003530:Cyprinidae / Q000378:metabolism*; D004958:Estradiol / Q000737:chemistry; D017930:Genes, Reporter; D057166:High-Throughput Screening Assays / Q000379:methods*; D008156:Luciferases / Q000032:analysis* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D020798:Two-Hybrid System Techniques
C485195:luciferase, cypridina; D004958:Estradiol; D008156:Luciferases