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US 7,622,275 B2Granted
Aequorin as a growth marker in yeast
AssigneeSanofi-Aventis Deutschland GmbH
InventorsPauline Fraissignes, Denis Guedin
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Abstract
A yeast cell containing constitutively expressed aequorin and methods of using the cells in growth and toxicity assays are disclosed.
Claims2
A method of measuring the growth rate of a yeast cell culture, the method comprising:
- (a) providing a yeast cell culture wherein the yeast cell comprises an aequorin-encoding deoxyribonucleic acid sequence expressibly linked to a constitutive yeast promoter, said constitutive yeast promoter being a triosephosphate isomerase promoter,
- (b) incubating the yeast cell culture and
- (c) measuring the amount of aequorin produced after the cells are lysed and mixed with Ca2+ by measuring luminescence wherein the intensity of luminescence indicates the growth rate of said yeast cell culture and wherein the intensity of luminescence is proportional to the growth rate at a cell density of up to 4×106 cells/ml.
A method for determining growth-regulating or toxic effects of a compound on a yeast cell culture, the method comprising:
- (a) providing a yeast cell comprising an aequorin-encoding deoxyribonucleic acid sequence expressibly linked to a constitutive yeast promoter, said constitutive yeast promoter being a triosephosphate isomerase promoter,
- (b) incubating the yeast cell with the compound,
- (c) measuring the amount of aequorin produced after the cells are lysed and mixed with Ca2+ by measuring luminescence wherein the intensity of luminescence indicates the growth rate of said yeast cell culture and wherein the intensity of luminescence is proportional to the growth rate at a cell density of up to 4×106 cells/ml and
- (d) comparing the measured amount of aequorin produced with that amount of aequorin produced by a yeast cell of step (a) incubated in the absence of the compound.
1 more element
Dependencies are inferred from the claim text.