Method for Verifying and/or Identifying Hormonally Effective Substances
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Abstract
The invention relates to a method for verifying and/or identifying hormonally effective substances using the pheromone system of yeast, to yeast cells for carrying out the method, and to the use thereof. The method according to the invention comprises the following steps: a) adding yeast culture medium or buffer to a sample which presumably contains a hormonally effective substance, b) thereafter contacting the sample with haploid yeast cells that possess features 1 through 3: 1. the DNA sequence coded for a heterologous hormone receptor is placed under the control of a constitutive promoter, 2. the DNA sequence coded for a yeast pheromone is placed under the control of a promoter that can be regulated by the hormone receptor, 3. a gene that is coded for a receptor for the yeast pheromone is constitutively expressed, wherein the activation of the hormone receptor by the hormonally effective substance leads to the expression and secretion of the yeast pheromone, and c) the physiological and/or morphological changes of haploid yeast cells are detected.
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Citations
62 Claims
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1-26. -26. (canceled)
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27. A method for verifying and/or identifying hormonally effective substances, comprising the steps of:
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a) to a sample that comprises presumably a hormonally active substance a yeast culturing medium or buffer is added, b) the sample to which the yeast culturing medium or buffer has been added is contacted with haploid yeast cells that comprise the following features 1 to 3, wherein either all features 1 to 3 are realized by a single type of haploid yeast cells or the features 1 and 2 are realized by a first type of haploid yeast cells and the feature 3 by a second type of haploid yeast cells; 1. the DNA sequence that codes for a heterologous hormone receptor is subjected to the control of a constitutive promoter and is functionally expressed; 2. the DNA sequence coding for a yeast pheromone is subjected to the control of a promoter that can be regulated by the heterologous hormone receptor; 3. a gene that codes for a receptor for the yeast pheromone is constitutively and functionally expressed,
wherein the activation of the heterologous hormone receptor by the hormonally active substance causes the expression of the DNA sequence coding for the yeast pheromone and the secretion of the pheromone from the haploid yeast cell; andc) a morphological change and/or a physiological change of the haploid yeast cells are detected. - View Dependent Claims (28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 44, 45, 46, 47, 48, 49, 50)
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43. The method according to 42 for detecting doping agents in urine samples.
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51. Haploid yeast cells with the following features:
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a DNA sequence that codes for a heterologous hormone receptor is subjected to the control of a constitutive promoter and is functionally expressed; a DNA sequence that codes for a yeast pheromone is subjected to the control of a promoter that can be regulated by the heterologous hormone receptor; wherein the activation of the heterologous hormone receptor by the hormonally active substance causes the expression of the DNA sequence coding for the yeast pheromone and the secretion of the yeast pheromone from the haploid yeast cell. - View Dependent Claims (52, 53, 54, 56, 57, 58, 59, 60, 61, 62)
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55. The haploid yeast cells according to 53 for detecting doping agents in urine samples.
Specification