An improved bioluminescence-based signaling assay for odor sensing with a yeast expressing a chimeric olfactory receptor.
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.
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Tokyo, Japan.
Yosuke
Y
Fukutani
Jun
J
Ishii
Keiichi
K
Noguchi
Akihiko
A
Kondo
Masafumi
M
Yohda
D001709:Biotechnology / Q000379:methods*; D004136:Dinitrobenzenes / Q000032:analysis; D044385:GTP-Binding Protein alpha Subunits / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D049409:Luciferases, Firefly / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D009812:Odorants / Q000032:analysis*; D011401:Promoter Regions, Genetic; D018035:Receptors, Odorant / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
D004136:Dinitrobenzenes; D044385:GTP-Binding Protein alpha Subunits; D018035:Receptors, Odorant; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; C016403:2,4-dinitrotoluene; D049409:Luciferases, Firefly