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An improved bioluminescence‐based signaling assay for odor sensing with a yeast expressing a chimeric olfactory receptor
Author(s) -
Fukutani Yosuke,
Ishii Jun,
Noguchi Keiichi,
Kondo Akihiko,
Yohda Masafumi
Publication year - 2012
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.24589
Subject(s) - bioluminescence , yeast , luciferase , g protein coupled receptor , reporter gene , biology , receptor , olfactory receptor , saccharomyces cerevisiae , microbiology and biotechnology , biochemistry , gene , transfection , gene expression
Abstract The goal of this work was to improve the bioluminescence‐based signaling assay system to create a practical application of a biomimetic odor sensor using an engineered yeast‐expressing olfactory receptors (ORs). Using the yeast endogenous pheromone receptor (Ste2p) as a model GPCR, we determined the suitable promoters for the firefly luciferase ( luc ) reporter and GPCR genes. Additionally, we deleted some genes to further improve the sensitivity of the luc reporter assay. By replacing the endogenous yeast G‐protein α‐subunit (Gpa1p) with the olfactory‐specific Gα olf , the optimized yeast strain successfully transduced signal through both OR and yeast Ste2p. Our results will assist the development of a bioluminescence‐based odor‐sensing system using OR‐expressing yeast. Biotechnol. Bioeng. 2012; 109: 3143–3151. © 2012 Wiley Periodicals, Inc.