Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Tokyo, Japan.
Biotechnol Bioeng. 2012 Dec;109(12):3143-51. doi: 10.1002/bit.24589. Epub 2012 Jul 4.
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.
本工作旨在改进基于生物发光的信号检测系统，利用工程化酵母表达嗅觉受体（OR），创建仿生气味传感器的实际应用。我们以酵母内源信息素受体（Ste2p）作为模型 GPCR，确定了萤火虫荧光素酶（luc）报告基因和 GPCR 基因的合适启动子。此外，我们删除了一些基因，以进一步提高 luc 报告基因检测的灵敏度。通过用嗅觉特异性 Gα（olf）替换内源性酵母 G 蛋白 α 亚基（Gpa1p），优化后的酵母菌株成功地通过 OR 和酵母 Ste2p 转导信号。我们的结果将有助于开发基于 OR 表达酵母的生物发光气味传感系统。
