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US4601978A
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Mammalian metallothionein promoter system
Assignee: University of California
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Filing: 1982-11-24
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Granted: 1986-07-22
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Expires: 2002-11-24
Abstract
Abstract A cloning vector comprises a replication system from a bovine papilloma virus, a regulatory system including the promoter and terminator from a human metallothionein II gene, and a prokaryotic replication system. The vector is useful for transforming mammalian cells where it is maintained as an automonously replicating episome. Transcription may be regulated by the presence of heavy metals and/or glucocorticoid hormones in the growth media.
Inventors
- 👤 Michael Karin
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 18:01
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85; C12N15/86; C12N2710/00; C12N2710/00011; C12N2710/20011; C12N2710/20041; C12N2710/20043; C12N2830/00; C12N2830/001; C12N2830/002; C12N2830/80; C12N2830/85
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
25 words
A DNA sequence of less than 500 base pairs, said DNA sequence comprising the human MT-II transcriptional regulatory system further comprising the transcription initiation sequence.
Claim 2
Dependent Claim
25 words
A DNA sequence of less than 500 base pairs, said DNA sequence comprising the human MT-II transcriptional regulatory system further comprising the transcription initiation sequence.
Claim 3
Dependent Claim
27 words
A DNA sequence as in claim 1, further comprising the human MT-II translation initiation codon located downstream from the transcription initiation sequence in the direction of transcription.
Claim 4
Dependent Claim
37 words
A DNA sequence as in claim 1, further comprising the human MT-II transcription termination sequence downstream from the transcription initiation sequence and at least one restriction site intermediate said transcription initiation sequence and said transcription termination sequence.
Claim 5
Dependent Claim
21 words
A DNA sequence as in claim 3, further comprising a structural gene other than human MT-II inserted at said restriction site.
Claim 6
Dependent Claim
67 words
A DNA construct capable of regulated expression of an inserted gene in a mammalian host, said construct comprising an extrachromosomal replication system recognized by a mammalian host, which replication system has been joined to a human MT-II gene inducible regulatory system in vitro, said construct having DNA sequence(s) defining at least one restriction site within the transcriptional control of the regulatory region for insertion of the gene.
Claim 7
Dependent Claim
67 words
A DNA construct capable of regulated expression of an inserted gene in a mammalian host, said construct comprising an extrachromosomal replication system recognized by a mammalian host, which replication system has been joined to a human MT-II gene inducible regulatory system in vitro, said construct having DNA sequence(s) defining at least one restriction site within the transcriptional control of the regulatory region for insertion of the gene.
Claim 8
Dependent Claim
18 words
A DNA construct as in claim 5, wherein the mammalian extrachromosomal replication system is derived from a papovavirus.
Claim 9
Dependent Claim
14 words
A DNA construct as in claim 6, wherein the papovavirus is bovine papilloma virus.
Claim 10
Dependent Claim
13 words
A DNA construct as in claim 5, further comprising a prokaryotic replication system.
Claim 11
Dependent Claim
16 words
A DNA construct as in claim 8, wherein the prokaryotic replication system is derived from pBR322.
Claim 12
Dependent Claim
60 words
A DNA construct useful for expression of a structural gene to produce a polypeptide in a mammalian host, said vector comprising (a) a replicon from bovine papilloma virus and (b) a regulatory system from a human MT-II gene comprising a promoter and a terminator and having at least one restriction site downstream from the promoter in the direction of transcription.
Claim 13
Dependent Claim
60 words
A DNA construct useful for expression of a structural gene to produce a polypeptide in a mammalian host, said vector comprising (a) a replicon from bovine papilloma virus and (b) a regulatory system from a human MT-II gene comprising a promoter and a terminator and having at least one restriction site downstream from the promoter in the direction of transcription.
Claim 14
Dependent Claim
31 words
A DNA construct as in claim 10, wherein the regulatory system is derived from the human MT-II gene and the restriction site is in the structural region of said MT-II gene.
Claim 15
Dependent Claim
34 words
A DNA construct as in claim 10, further comprising a structural gene inserted into the restriction site so that expression of the structural gene is under the regulatory control of the promoter and terminator.
Claim 16
Dependent Claim
14 words
A mammalian cell transformed with at least one DNA construct according to claim 12.
Claim 17
Dependent Claim
15 words
A mouse fibroblast cell transformed with at least one DNA construct according to claim 12.
Claim 18
Dependent Claim
59 words
A method for producing a polypeptide encoded by a structural gene, said method comprising: transforming a mammalian cell with a DNA construct according to claim 12; growing the transformed cell in an appropriate nutrient medium; inducing expression of the structural gene by the addition of a heavy metal or a glucocorticoid to the nutrient medium; and isolating the polypeptide.
Claim 19
Dependent Claim
14 words
A method as in claim 15, wherein the mammalian cell is a mouse fibroblast.
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) | 1989-07-21 | 1990-01-20 | 1990-07-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-07-21 | 1994-01-20 | 1994-07-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-07-21 | 1998-01-20 | 1998-07-22 | $7700.00 | Window Closed / Lapsed |