Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
208 words
A bioassay for evaluating compounds as potential antagonists for members of the steroid hormone superfamily of receptor protein(s), said bioassay comprising: (a) culturing test cells in culture medium containing: increasing concentrations of at least one compound whose ability to inhibit the action of agonists on members of the steroid hormone superfamily of receptors is sought to be determined, and a fixed concentration of at least one agonist for said hormone receptor protein(s), wherein said test cells contain: DNA which expresses a member of the steroid hormone superfamily of receptor protein(s), in a form having improved trans-activation transcription activity, relative to wild-type receptor wherein said member contains a plurality of at least one parental receptor trans-activation domain selected from τ 1 or τ 2 , located outside the DNA-binding and ligand-binding domain(s) of said receptor protein(s), and a DNA sequence comprising a response element for said hormone receptor protein, operatively linked to a reporter gene; and thereafter (b) monitoring in said cells the level of expression of the product of said reporter gene as a function of the concentration of said potential antagonist compound in said culture medium, thereby indicating the ability of said potential antagonist compound to inhibit activation of members of the steroid hormone superfamily of receptors.
Claim 2
Dependent Claim
208 words
A bioassay for evaluating compounds as potential antagonists for members of the steroid hormone superfamily of receptor protein(s), said bioassay comprising: (a) culturing test cells in culture medium containing: increasing concentrations of at least one compound whose ability to inhibit the action of agonists on members of the steroid hormone superfamily of receptors is sought to be determined, and a fixed concentration of at least one agonist for said hormone receptor protein(s), wherein said test cells contain: DNA which expresses a member of the steroid hormone superfamily of receptor protein(s), in a form having improved trans-activation transcription activity, relative to wild-type receptor wherein said member contains a plurality of at least one parental receptor trans-activation domain selected from τ 1 or τ 2 , located outside the DNA-binding and ligand-binding domain(s) of said receptor protein(s), and a DNA sequence comprising a response element for said hormone receptor protein, operatively linked to a reporter gene; and thereafter (b) monitoring in said cells the level of expression of the product of said reporter gene as a function of the concentration of said potential antagonist compound in said culture medium, thereby indicating the ability of said potential antagonist compound to inhibit activation of members of the steroid hormone superfamily of receptors.
Claim 3
Dependent Claim
17 words
A bioassay according to claim 1 wherein said receptor protein contains two of said τ 2 domains.
Claim 4
Dependent Claim
20 words
A bioassay according to claim 2 wherein the second of said τ 2 domains is located adjacent to the first.
Claim 5
Dependent Claim
23 words
A bioassay according to claim 2 wherein the second of said τ 2 domains is located distal from the first τ 2 sequence.
Claim 6
Dependent Claim
17 words
A bioassay according to claim 1 wherein said receptor protein contains two of said τ 1 domains.
Claim 7
Dependent Claim
20 words
A bioassay according to claim 5 wherein the second of said τ 1 domains is located adjacent to the first.
Claim 8
Dependent Claim
23 words
A bioassay according to claim 5 wherein the second of said τ 1 domains is located distal from the first τ 1 sequence.
Claim 9
Dependent Claim
17 words
A bioassay according to claim 1 wherein said host cells are selected from CV-1 or COS cells.
Claim 10
Dependent Claim
20 words
A bioassay according to claim 1 wherein said reporter and expression plasmids also contain the origin of replication of SV-40.
Claim 11
Dependent Claim
17 words
A bioassay according to claim 1 wherein said reporter and expression plasmids also contain a selectable marker.
Claim 12
Dependent Claim
33 words
A bioassay according to claim 1 wherein said hormone response element is selected from mouse tumor virus long terminal repeat (MTV LTR) or mammalian growth hormone promoter, and said reporter gene is CAT.
Claim 13
Dependent Claim
16 words
A bioassay according to claim 1 wherein said receptor protein(s) is selected from steroid hormone receptors.