Patent US4559302A successfully refreshed from Google Patents.
US4559302A
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DNA for directing transcription and expression of structural genes
Assignee: Eli Lilly and Co
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Filing: 1982-11-01
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Granted: 1985-12-17
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Expires: 2002-11-01
Abstract
Abstract The present invention relates to a novel transcriptional and translational activating sequence. The novel activating sequence can be either chemically synthesized or isolated on a 0.17 kb PstI-SacI restriction fragment from plasmid pKC203, a plasmid of E. coli JR225 (ATCC 31912). The activating sequence directs expression of the aminoglycoside acetyltransferase aac(3)IV and hygromycin phosphotransferase aph(4) genes present on plasmid pKC203. A series of expression vectors have been constructed in which the activating sequence directs the expression of beta-galactosidase or hygromycin phosphotransferase. These vectors can be readily modified and have been designed to facilitate the subsequent cloning and expression of any gene of research or commercial interest. The expression and cloning vectors have been transformed into E. coli and other host cells in which the activating sequence functions.
Inventors
- 👤 Thomas D. Ingolia
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 08:15
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/70; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/848; Y10S435/849; Y10S435/886
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
24 words
A constructed DNA consisting essentially of the promoter, Shine-Dalgarno sequence, and translational start triplet of the polycistronic apramycin (aac(3)IV) and hygromycin (aph(4)) resistance genes.
Claim 2
Dependent Claim
24 words
A constructed DNA consisting essentially of the promoter, Shine-Dalgarno sequence, and translational start triplet of the polycistronic apramycin (aac(3)IV) and hygromycin (aph(4)) resistance genes.
Claim 3
Dependent Claim
22 words
A constructed DNA which comprises the sequence ##STR2## wherein A is deoxyadenyl, G is deoxyguanyl, C is deoxycytosyl and T is thymidyl.
Claim 4
Dependent Claim
22 words
A constructed DNA which comprises the sequence ##STR2## wherein A is deoxyadenyl, G is deoxyguanyl, C is deoxycytosyl and T is thymidyl.
Claim 5
Dependent Claim
49 words
A recombinant DNA cloning vector which comprises the sequence ##STR3## wherein A is deoxyadenyl, G is deoxyguanyl, C is deoxycytosyl, T is thymidyl, and R is a structural gene, exclusive of the translational start triplet, that codes for a functional polypeptide that is heterologous with respect to E. coli.
Claim 6
Dependent Claim
49 words
A recombinant DNA cloning vector which comprises the sequence ##STR3## wherein A is deoxyadenyl, G is deoxyguanyl, C is deoxycytosyl, T is thymidyl, and R is a structural gene, exclusive of the translational start triplet, that codes for a functional polypeptide that is heterologous with respect to E. coli.
Claim 7
Dependent Claim
12 words
The recombinant DNA cloning vector of claim 3 which is a plasmid.
Claim 8
Dependent Claim
36 words
A DNA restriction fragment which is selected from the group consisting of the ˜0.17 kb PstI-SacI fragment of plasmid pKC222, ˜0.17 kb PstI-BamHI fragment of plasmid pTI104, and the ˜0.17 kb PstI-BamHI fragment of plasmid pTI106.
Claim 9
Dependent Claim
36 words
A DNA restriction fragment which is selected from the group consisting of the ˜0.17 kb PstI-SacI fragment of plasmid pKC222, ˜0.17 kb PstI-BamHI fragment of plasmid pTI104, and the ˜0.17 kb PstI-BamHI fragment of plasmid pTI106.
Claim 10
Dependent Claim
51 words
The plasmid of claim 4 in which the structural gene is selected from the group consisting of genes that code for β-galactosidase, human pre-proinsulin, human proinsulin, human insulin A-chain, human insulin β-chain, non-human insulin, human growth hormone, non-human growth hormone, bovine growth hormone, human interferon, non-human interferon, viral antigen, and urokinase.
Claim 11
Dependent Claim
13 words
The plasmid of claim 6 in which the structural gene codes for β-galactosidase.
Claim 12
Dependent Claim
14 words
The plasmid of claim 6 in which the structural gene codes for human proinsulin.
Claim 13
Dependent Claim
15 words
The plasmid of claim 6 in which the structural gene codes for human insulin A-chain.
Claim 14
Dependent Claim
15 words
The plasmid of claim 6 in which the structural gene codes for human growth hormone.
Claim 15
Dependent Claim
15 words
The plasmid of claim 6 in which the structural gene codes for bovine growth hormone.
Claim 16
Dependent Claim
15 words
The plasmid of claim 6 in which the structural gene codes for human insulin β-chain.
Claim 17
Dependent Claim
17 words
The plasmid of claim 6 which is selected from the group consisting of plasmid pTI104 and pTI106.
Claim 18
Dependent Claim
55 words
A bacterial host cell which is transformed by a recombinant DNA cloning vector which comprises ##STR4## wherein A is deoxyadenyl, G is deoxyguanyl, C is deoxycytosyl, T is thymidyl, and R is a structural gene, exclusive of the translational start triplet, that codes for a functional polypeptide that is heterologous with respect to E. coli.
Claim 19
Dependent Claim
55 words
A bacterial host cell which is transformed by a recombinant DNA cloning vector which comprises ##STR4## wherein A is deoxyadenyl, G is deoxyguanyl, C is deoxycytosyl, T is thymidyl, and R is a structural gene, exclusive of the translational start triplet, that codes for a functional polypeptide that is heterologous with respect to E. coli.
Claim 20
Dependent Claim
16 words
The host cell of claim 14 in which the recombinant DNA cloning vector is a plasmid.
Claim 21
Dependent Claim
10 words
The host cell of claim 14 which is E. coli.
Claim 22
Dependent Claim
43 words
The host cell of claim 16 which is selected from the group consisting of E. coli K12 RRl/pTI104, E. coli K12 RRl/PTI106, E. coli K12 JA221/pTI106, E. coli K12 JA221/pTI104, E. coli K12 HB101/pTI104, E. coli K12 HB101/pTI106 and E. coli K12 BE904/pTI106.
Claim 23
Dependent Claim
11 words
The host cell of claim 15 which is a Streptomyces cell.
References Cited (Backward Citations)
US4237224A
Cited
US4349629A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1988-12-16 | 1989-06-17 | 1989-12-17 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-12-16 | 1993-06-17 | 1993-12-17 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-12-16 | 1997-06-17 | 1997-12-17 | $7700.00 | Window Closed / Lapsed |