Functional expression of olfactory receptors in yeast and development of a bioassay for odorant screening

作者: Jasmina Minic , Marie-Annick Persuy , Elodie Godel , Josiane Aioun , Ian Connerton

DOI: 10.1111/J.1742-4658.2004.04494.X

关键词:

摘要: The functional expression of olfactory receptors (ORs) is a primary requirement to examine the molecular mechanisms odorant perception and coding. Functional rat I7 OR its trafficking plasma membrane was achieved under optimized experimental conditions in budding yeast Saccharomyces cerevisiae. receptor shown by Western blotting immunolocalization methods. Moreover, we took advantage similarities between signal transduction cascades G protein-coupled mammalian cells pheromone response pathway develop novel biosensor for screening using luciferase as reporter. Yeasts were engineered coexpress Gα subunit, compensate lack endogenous Gpa1 so that stimulation ligands activates MAP kinase signaling induces synthesis. sensitivity bioassay significantly enhanced Golf compared Gα15 resulting dose-dependent responses system. probed with an array odorants demonstrate yeast-borne retains specificity selectivity towards ligands. results are confirmed bioluminescence human OR17-40 specific ligand, helional. Based on these findings, reporter should be amenable simple, rapid inexpensive hundreds ORs provide insight into coding mechanisms.

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