US5422262A
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Steroid 5a reductase nucleic acid segments and recombinant vectors and host cells
Assignee: The University of Texas System
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Filing: 1991-11-18
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Granted: 1995-06-06
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Expires: 2011-11-18
Abstract
Abstract Disclosed are methods and compositions for the preparation of steroid 5 alpha -reductases by recombinant means, as well as for the use of these enzymes in screening assays for the identification of compounds which have the ability to inhibit or otherwise alter the enzymatic function of these enzymes. Biochemical and pharmacological evidence is presented to demonstrate the existence of more than one human steroid 5 alpha -reductase. The DNA sequence encoding steroid 5 alpha -reductase 2, the major active isozyme of human genital tissue, is disclosed herein, in addition to methods and compositions for its preparation and pharmacological analysis. The sequences disclosed herein may be used directly in the preparation of genetic constructs, or may be employed in the preparation of hybridization probes for the selection of enzyme-encoding sequences from other sources. These sequences may prove useful in an analysis of normal and abnormal sexual differentiation, benign prostatic hyperplasia, male pattern baldness, acne, hirsutism, endometriosis, and cancer of the prostate.
Inventors
- 👤 Sefan Andersson
- 👤 David W. Russell
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 10:13
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/001; C12Q; C12Q1/00; C12Q1/26; C12Q1/32; C12Y; C12Y103/00; C12Y103/01; C12Y103/01022
Claims Analysis
Total Claims: 49
Claim 1
Independent Claim
52 words
A purified and isolated recombinant DNA segment comprising a coding region encoding a Type 1 rat or Type 1 human steroid 5α-reductase, the coding region encoding an amino acid sequence consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:4, or the full complement of such a region.
Claim 2
Dependent Claim
52 words
A purified and isolated recombinant DNA segment comprising a coding region encoding a Type 1 rat or Type 1 human steroid 5α-reductase, the coding region encoding an amino acid sequence consisting of the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:4, or the full complement of such a region.
Claim 3
Dependent Claim
34 words
The nucleic acid segment of claim 1, wherein the encoded steroid 5α-reductase consists of an amino acid sequence that set forth in SEQ ID NO:4, or the full complement of such a coding region.
Claim 4
Dependent Claim
38 words
The nucleic acid segment of claim 2, wherein the coding region is further defined as consisting of the nucleic acid sequence coding region set forth in SEQ ID NO:3, or the full complement of such a coding region.
Claim 5
Dependent Claim
42 words
The nucleic acid segment of claim 1, wherein the segment comprises a steroid 5α-reductase coding region that encodes a steroid 5α-reductase amino acid sequence consisting of that set forth in SEQ ID NO:2, or the full complement of such a coding sequence.
Claim 6
Dependent Claim
38 words
The nucleic acid segment of claim 4, wherein the segment comprises a steroid 5α-reductase-encoding nucleic acid sequence consisting of the coding region set forth in SEQ ID NO: 1, or the full complement of such a coding region.
Claim 7
Dependent Claim
38 words
A recombinant vector which comprises a DNA segment encoding a Type 1 rat or Type 1 human steroid 5α-reductase enzyme having an amino acid sequence consisting of that set forth in SEQ ID NO:2, or SEQ ID NO:4.
Claim 8
Dependent Claim
38 words
A recombinant vector which comprises a DNA segment encoding a Type 1 rat or Type 1 human steroid 5α-reductase enzyme having an amino acid sequence consisting of that set forth in SEQ ID NO:2, or SEQ ID NO:4.
Claim 9
Dependent Claim
45 words
A purified and isolated recombinant acid segment comprising a region which consists of: (a) at least a thirty nucleotide long contiguous stretch of the nucleic acid sequence shown in SEQ ID NO: 1 or SEQ ID NO:3; or (b) the full complement of said sequence.
Claim 10
Dependent Claim
45 words
A purified and isolated recombinant acid segment comprising a region which consists of: (a) at least a thirty nucleotide long contiguous stretch of the nucleic acid sequence shown in SEQ ID NO: 1 or SEQ ID NO:3; or (b) the full complement of said sequence.
Claim 11
Dependent Claim
47 words
The nucleic acid segment of claim 7, wherein the segment comprises a region that consists of: (a) at least a forty nucleotide long contiguous stretch of the nucleic acid sequence shown in SEQ ID NO: 1 or SEQ ID NO:3; (b) the full complement of said sequence.
Claim 12
Dependent Claim
21 words
The nucleic acid segment of claim 7, further defined as comprising a DNA fragment of up to 200 basepairs in length.
Claim 13
Dependent Claim
21 words
The nucleic acid segment of claim 9, further defined as comprising a DNA fragment of up to 100 basepairs in length.
Claim 14
Dependent Claim
21 words
The nucleic acid segment of claim 10, further defined as comprising a DNA fragment of up to 50 basepairs in length.
Claim 15
Dependent Claim
29 words
The vector of claim 6, wherein the vector encodes human Type 1 steroid 5α-reductase, as defined by the amino acid sequence consisting of that shown in SEQ ID NO:4.
Claim 16
Dependent Claim
25 words
The vector of claim 12, wherein the encoded steroid 5α-reductase comprises asteroid 5α-reductase amino acid sequence consisting of that set forth in SEQ ID NO:4.
Claim 17
Dependent Claim
28 words
The vector of claim 12, wherein the encoded Type 1 rat steroid 5α-reductase comprises asteroid 5α-reductase amino acid sequence consisting of that set forth in SEQ ID NO:2.
Claim 18
Dependent Claim
25 words
The vector of claim 14, further defined as comprising a steroid 5α-reductase-encoding nucleic acid sequence consisting of that set forth in SEQ ID NO: 1.
Claim 19
Dependent Claim
39 words
A vector comprising a DNA segment, the DNA segment consisting of at least a fifteen thirty nucleotide long contiguous stretch of a DNA sequence of SEQ ID NO:1, SEQ ID NO:3 or SEQ.ID NOS:7, 9-11, 13-15, 17-19, and 21-23.
Claim 20
Dependent Claim
39 words
A vector comprising a DNA segment, the DNA segment consisting of at least a fifteen thirty nucleotide long contiguous stretch of a DNA sequence of SEQ ID NO:1, SEQ ID NO:3 or SEQ.ID NOS:7, 9-11, 13-15, 17-19, and 21-23.
Claim 21
Dependent Claim
39 words
The vector of claim 16, wherein the segment consists of at least a forty nucleotide long contiguous stretch of the DNA sequence of SEQ ID NO: 1, SEQ ID NO:3 or SEQ ID NOS:7, 9-11, 13-15, 17-19, and 21-23.
Claim 22
Dependent Claim
19 words
The vector of claim 16, further defined as comprising a DNA fragment of up to 200 basepairs in length.
Claim 23
Dependent Claim
19 words
The vector of claim 18, further defined as comprising a DNA fragment of up to 100 basepairs in length.
Claim 24
Dependent Claim
19 words
The vector of claim 19, further defined as comprising a DNA fragment of up to 50 basepairs in length.
Claim 25
Dependent Claim
40 words
The vector of claim 16, wherein the segment encodes a Type 1 rat or Type 1 human steroid 5α-reductase enzyme, as defined by the amino acid sequence consisting of that set forth in SEQ ID NO:2 or SEQ ID NO:4.
Claim 26
Dependent Claim
22 words
A recombinant host cell incorporating a recombinant vector in accordance with any one of claims 12, 13, 14, 16 or 17-21 .
Claim 27
Dependent Claim
48 words
The recombinant host cell of claim 22, wherein the recombinant DNA segment is further defined as one encoding a Type 1 rat or Type 1 human steroid 5α-reductase enzyme, as defined by the amino acid sequence consisting of that shown in SEQ ID NO:2 or SEQ ID NO:4.
Claim 28
Dependent Claim
14 words
The recombinant host cell of claim 22, further defined as a eukaryotic host cell.
Claim 29
Dependent Claim
20 words
The recombinant host cell of claim 24, wherein the DNA segment is integrated into the genome of the host cell.
Claim 30
Dependent Claim
17 words
The recombinant host cell of claim 22, wherein the DNA segment is positioned on a recombinant vector.
Claim 31
Dependent Claim
34 words
The vector of claim 16, wherein the segment consists of at least a thirty nucleotide long contiguous stretch of the coding sequence of the DNA sequence of SEQ ID NO:1 or SEQ ID NO:3.
Claim 32
Dependent Claim
34 words
The vector of claim 27 wherein the segment consists of at least a forty nucleotide long contiguous stretch of the coding sequence of the DNA sequence of SEQ ID NO:1 or SEQ ID NO:3.
Claim 33
Dependent Claim
45 words
A purified and isolated recombinant nucleic acid segment comprising a coding region encoding human Type 2 steroid 5α-reductase, the coding region encoding an amino acid sequence consisting of the amino acid sequence of SEQ ID NO:6, or the full complement of such a coding region.
Claim 34
Dependent Claim
45 words
A purified and isolated recombinant nucleic acid segment comprising a coding region encoding human Type 2 steroid 5α-reductase, the coding region encoding an amino acid sequence consisting of the amino acid sequence of SEQ ID NO:6, or the full complement of such a coding region.
Claim 35
Dependent Claim
34 words
The nucleic acid segment of claim 29, further defined as comprising asteroid 5α-reductase-encoding nucleic acid sequence consisting of that set forth in SEQ ID NO:5, or the full complement of such a coding region.
Claim 36
Dependent Claim
44 words
A purified and isolated recombinant nucleic acid segment comprising a region, the region consisting of at least a thirty nucleotide long contiguous stretch of the nucleic acid sequence of nucleotides 1 through 2437 of SEQ ID NO:5, or the full complement of said sequence.
Claim 37
Dependent Claim
44 words
A purified and isolated recombinant nucleic acid segment comprising a region, the region consisting of at least a thirty nucleotide long contiguous stretch of the nucleic acid sequence of nucleotides 1 through 2437 of SEQ ID NO:5, or the full complement of said sequence.
Claim 38
Dependent Claim
45 words
The nucleic acid segment of claim 31, wherein the segment comprises a region that consists of at least a forty nucleotide long contiguous stretch of the nucleic acid sequence of nucleotides 1 through 2437 of SEQ ID NO:5, or the full complement of said sequence.
Claim 39
Dependent Claim
19 words
The nucleic acid segment of claim 31, further defined as a fragment of up to 200 basepairs in length.
Claim 40
Dependent Claim
19 words
The nucleic acid segment of claim 33, further defined as a fragment of up to 100 basepairs in length.
Claim 41
Dependent Claim
19 words
The nucleic acid segment of claim 34, further defined as a fragment of up to 50 basepairs in length.
Claim 42
Dependent Claim
15 words
The nucleic acid segment of claims 29 or 31, further defined as a DNA segment.
Claim 43
Dependent Claim
15 words
The nucleic acid segment of claims 29 or 31, further defined as an RNA segment.
Claim 44
Dependent Claim
15 words
The nucleic acid segment of claims 29 or 31, further defined as a recombinant vector.
Claim 45
Dependent Claim
17 words
A recombinant host cell comprising a recombinant nucleic acid segment in accordance with claims 29 or 31.
Claim 46
Dependent Claim
14 words
The recombinant host cell of claim 39, further defined as a eukaryotic host cell.
Claim 47
Dependent Claim
20 words
The recombinant host cell of claim 39, wherein the DNA segment is integrated into the genome of the host cell.
Claim 48
Dependent Claim
17 words
The recombinant host cell of claim 39, wherein the DNA segment is positioned on a recombinant vector.
Claim 49
Dependent Claim
18 words
The recombinant host cell of claim 39, further defined as expressing asteroid 5α-reductase having steroid 5α-reductase enzymatic activity.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1998-06-05 | 1998-12-05 | 1999-06-06 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2002-06-05 | 2002-12-05 | 2003-06-06 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2006-06-05 | 2006-12-05 | 2007-06-06 | $7700.00 | Window Closed / Lapsed |