| Title: | An improved bioluminescence-based signaling assay for odor sensing with a yeast expressing a chimeric olfactory receptor |
| Author(s): | Fukutani Y; Ishii J; Noguchi K; Kondo A; Yohda M; |
| Address: | "Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Tokyo, Japan" |
| ISSN/ISBN: | 1097-0290 (Electronic) 0006-3592 (Linking) |
| 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 alpha-subunit (Gpa1p) with the olfactory-specific Galpha(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" |
| Keywords: | "Biotechnology/*methods Dinitrobenzenes/analysis GTP-Binding Protein alpha Subunits/genetics/metabolism Immunohistochemistry Luciferases, Firefly/chemistry/genetics/metabolism Luminescent Measurements/*methods Odorants/*analysis Promoter Regions, Genetic R;" |
| Notes: | "MedlineFukutani, Yosuke Ishii, Jun Noguchi, Keiichi Kondo, Akihiko Yohda, Masafumi eng Research Support, Non-U.S. Gov't 2012/06/26 Biotechnol Bioeng. 2012 Dec; 109(12):3143-51. doi: 10.1002/bit.24589. Epub 2012 Jul 4" |
