Yeast cells engineered to produce pheromone system protein surrogates, and uses therefor
First Claim
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1. A transformed yeast cell comprising a reporter gene under control of a pheromone-responsive promoter, a heterologous G protein-coupled receptor gene, each said gene being under the control of a separate promoter, a mutation in a SCG1/GPA1 gene, and a hybrid G α
- protein, which heterologous G protein coupled receptor gene encodes a receptor selected from the group consisting of a β
2 adrenergic receptor, an α
2-adrenergic receptor, a 5HT-1A receptor, a muscarinic acetylcholine receptor, a growth hormone releasing factor receptor and a somatostatin receptor.
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Abstract
Yeast cells are engineered to express both a surrogate of a pheromone system protein (e.g., enzymes involved in maturation of α-factor, transporters of a-factor, pheromone receptors, etc.) and a potential peptide modulator of the surrogate, in such a manner that the inhibition or activation of the surrogate affects a screenable or selectable trait of the yeast cells. Various additional features improve the signal-to-noise ratio of the screening/selection system.
12 Citations
2 Claims
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1. A transformed yeast cell comprising a reporter gene under control of a pheromone-responsive promoter, a heterologous G protein-coupled receptor gene, each said gene being under the control of a separate promoter, a mutation in a SCG1/GPA1 gene, and a hybrid G α
- protein, which heterologous G protein coupled receptor gene encodes a receptor selected from the group consisting of a β
2 adrenergic receptor, an α
2-adrenergic receptor, a 5HT-1A receptor, a muscarinic acetylcholine receptor, a growth hormone releasing factor receptor and a somatostatin receptor.
- protein, which heterologous G protein coupled receptor gene encodes a receptor selected from the group consisting of a β
-
2. A transformed yeast cell comprising a reporter gene under the control of a pheromone-responsive promoter, a gene encoding a heterologous G protein-coupled receptor, each said gene being under the control of a separate promoter, a mutation in SCG1/GPA1 gene, and a hybrid Gα
- protein.
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Current AssigneeCadus Technologies Incorporated
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Original AssigneeCadus Technologies Incorporated
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InventorsTrueheart, Joshua, Murphy, Andrew J., Klein, Christine, Paul, Jeremy, Fowlkes, Dana M., Broach, Jim, Manfredi, John
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Primary Examiner(s)Chernyshev; Olga N.
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Application NumberUS09/286,166Time in Patent Office3,431 DaysField of Search435/7.1, 435/7.2, 435/7.31, 435/7.8, 435/254.11, 435/255.1, 436/501US Class Current435/254.2CPC Class CodesC07K 14/395 from SaccharomycesC07K 2319/02 containing a signal sequenceC12N 15/81 for yeasts
