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US4766066A
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Method of using bacteriophage lambda p1 promoter to produce a functional polypeptide in streptomyces
Assignee: Eli Lilly and Co
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Filing: 1984-09-27
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Granted: 1988-08-23
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Expires: 2004-09-27
Abstract
Abstract The invention relates to a method and cloning vehicle for the expression of a functional polypeptide in Streptomyces. A recombinant DNA cloning vehicle was genetically engineered to bring the expression of the neomycin phosphotransferase gene under the control of the Escherichia coli bacteriophage λp L promoter.
Inventors
- 👤 Stuart A. Kuhstoss
- 👤 R. Nagaraja Rao
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 22:24
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/70; C12N15/73; C12N15/74; C12N15/76; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/886; Y10S435/889; Y10S435/896
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
115 words
A method of using bacteriophage lambda p L promoter to produce a functional polypeptide in Streptomyces, said method comprising: (a) transforming a Streptomyces host cell with a selectable and autonomously replicating recombinant DNA expression vector comprising: (1) the bacteriophage λp L promoter-operator transcriptional activating sequence; (2) a translational activating sequence; (3) a DNA sequence that codes for a functional polypeptide; and (b) culturing said transformed cell under conditions suitable for expression of said polypeptide, subject to the limitation that said expression vector sequentially contains said λp L promoter-operator transcriptional activating sequence, said translational activating sequence and said DNA sequence that codes for a functional polypeptide such that a translatable mRNA transcript encodes the functional polypeptide.
Claim 2
Independent Claim
115 words
A method of using bacteriophage lambda p L promoter to produce a functional polypeptide in Streptomyces, said method comprising: (a) transforming a Streptomyces host cell with a selectable and autonomously replicating recombinant DNA expression vector comprising: (1) the bacteriophage λp L promoter-operator transcriptional activating sequence; (2) a translational activating sequence; (3) a DNA sequence that codes for a functional polypeptide; and (b) culturing said transformed cell under conditions suitable for expression of said polypeptide, subject to the limitation that said expression vector sequentially contains said λp L promoter-operator transcriptional activating sequence, said translational activating sequence and said DNA sequence that codes for a functional polypeptide such that a translatable mRNA transcript encodes the functional polypeptide.
Claim 3
Dependent Claim
12 words
The method of claim 1 wherein the expression vector is a plasmid.
Claim 4
Dependent Claim
12 words
The method of claim 1 wherein the expression vector is plasmid pKC417.
Claim 5
Dependent Claim
34 words
The method of claim 1 in which the transformed Streptomyces host cell is selected from the group consisting of Streptomyces ambofaciens, Streptomyces aureofaciens, Streptomyces cinnamonesis, Streptomyces fradiae, Streptomyces qranuloruber, Streptomyces qriseofuscus, and Streptomyces lividans.
Claim 6
Dependent Claim
15 words
The method of claim 4 in which the transformed Streptomyces host cell is Streptomyces ambofaciens.
Claim 7
Dependent Claim
15 words
The method of claim 4 in which the transformed Streptomyces host cell is Streptomyces fradiae.
Claim 8
Dependent Claim
15 words
The method of claim 4 in which the transformed Streptomyces host cell is Streptomyces lividans.
Claim 9
Dependent Claim
63 words
The method of claim 1 wherein the gene that encodes a functional polypeptide is selected from the group consisting of genes that encode neomycin phosphotransferase, human pre-proinsulin, human proinsulin, human insulin A-chain, human insulin B-chain, non-human insulin, human growth hormone, non-human growth hormone, bovine growth hormone, porcine growth hormone, human interferon, non-human interferon, viral antigen, urokinase, any peptide hormone and any polypeptide enzyme.
Claim 10
Dependent Claim
18 words
The method of claim 8 wherein the DNA sequence that codes for a functional polypeptide is neomycin phosphotransferase.
Claim 11
Dependent Claim
19 words
The method of claim 1 wherein the bacteriophage λp L promoter-operator transcriptional activating sequence is derived from plasmid pKC283.
Claim 12
Dependent Claim
14 words
A plasmid selected from the group consisting of pKC309, pKC322, pKC283, pKC354, and pKC356.
Claim 13
Dependent Claim
14 words
A plasmid selected from the group consisting of pKC309, pKC322, pKC283, pKC354, and pKC356.
Claim 14
Dependent Claim
8 words
The plasmid of claim 11 which is pKC283.
Claim 15
Dependent Claim
91 words
A Streptomyces host cell transformed by a recombinant DNA expression vector capable of selection and autonomous replication in said Streptomyces host cell, said vector comprising: (a) the bacteriophage λP L promoter-operator transcription activating sequence; (b) a translational activating sequence; and (c) a DNA sequence that codes for a functional polypetide, subject to the limitation that said expression vector sequentially contains said λP L promoter-operator transcriptional activating sequence, said translational activating sequence, and said DNA sequence that codes for a functional polypeptide such that a translatable mRNA transcript encodes said functional polypeptide.
Claim 16
Dependent Claim
91 words
A Streptomyces host cell transformed by a recombinant DNA expression vector capable of selection and autonomous replication in said Streptomyces host cell, said vector comprising: (a) the bacteriophage λP L promoter-operator transcription activating sequence; (b) a translational activating sequence; and (c) a DNA sequence that codes for a functional polypetide, subject to the limitation that said expression vector sequentially contains said λP L promoter-operator transcriptional activating sequence, said translational activating sequence, and said DNA sequence that codes for a functional polypeptide such that a translatable mRNA transcript encodes said functional polypeptide.
Claim 17
Dependent Claim
12 words
The transformed Streptomyces host cell of claim 13 which is Streptomyces ambofaciens/pKC417.
Claim 18
Dependent Claim
83 words
A selectable recombinant DNA expression vector which replicates in Streptomyces which comprises: (a) the bacteriophage λP L promoter-operator transcriptional activating sequence; (b) a translational activating sequence; and (c) a DNA sequence that codes for a functional polypeptide, subject to the limitation that in a Streptomyces host cell said expression vector sequentially contains said λP L promoter-operater transcriptional activating sequence, said translational activating sequence, and said DNA sequence that codes for a functional polypeptide such that a translatable mRNA transcript encodes said functional polypeptide.
Claim 19
Dependent Claim
83 words
A selectable recombinant DNA expression vector which replicates in Streptomyces which comprises: (a) the bacteriophage λP L promoter-operator transcriptional activating sequence; (b) a translational activating sequence; and (c) a DNA sequence that codes for a functional polypeptide, subject to the limitation that in a Streptomyces host cell said expression vector sequentially contains said λP L promoter-operater transcriptional activating sequence, said translational activating sequence, and said DNA sequence that codes for a functional polypeptide such that a translatable mRNA transcript encodes said functional polypeptide.
Claim 20
Dependent Claim
9 words
The vector of claim 15 which is a plasmid.
Claim 21
Dependent Claim
8 words
The plasmid of claim 16 which is pKC417.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-08-23 | 1992-02-22 | 1992-08-23 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-08-23 | 1996-02-22 | 1996-08-23 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-08-23 | 2000-02-22 | 2000-08-23 | $7700.00 | Window Closed / Lapsed |