IP Library › Granted Patent US 7,622,275
Granted Patent B2
US 7,622,275 · App. 11/011,349 · Granted Nov 24, 2009

Aequorin as a growth marker in yeast

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Quick Facts
Patent No.
US 7,622,275
App. No.
11/011,349
Granted
Nov 24, 2009
Kind
B2
Abstract

A yeast cell containing constitutively expressed aequorin and methods of using the cells in growth and toxicity assays are disclosed.

Claims (9)

1. 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 Ca 2+ 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 4×10 6 cells/ml.

2. 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 Ca 2+ 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 4×10 6 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.