US4769327A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Secretion vector
Assignee: Omnigene Inc
•
Filing: 1986-03-28
•
Granted: 1988-09-06
•
Expires: 2006-03-28
Abstract
Abstract A vector including a DNA sequence encoding a secretory signal sequence substantially identical to the secretory signal encoding sequence of the Bacillus licheniformis alpha -amylase gene; upstream from the signal-encoding sequence, a promoter sequence and a ribosome binding site sequence, transcription of the signal-encoding sequence being under the control of the promoter sequence; and downstream from the signal-encoding sequence, a site for the insertion into the vector of a heterologous DNA sequence, in reading frame with the signal-encoding sequence.
Inventors
- 👤 Michael A. Stephens
- 👤 Cathy F. Rudolph
- 👤 Nancy M. Hannett
- 👤 Diane L. Stassi
- 👤 Janice G. Pero
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 14:11
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/87; C12N15/90; C07; C07K; C07K14/00; C07K14/435; C07K14/575; C07K14/58; C07K14/60; C12N15/63; C12N15/74; C12N15/75; C12N9/00; C12N9/14; C12N9/16; C12N9/24; C12N9/2402; C12N9/2405; C12N9/2408; C12N9/2411; C12N9/2414; C12N9/2417; C12N9/2428; C07K2319/00; C07K2319/01; C07K2319/02; C07K2319/036; C07K2319/70; C07K2319/74; C07K2319/75; Y; Y10; Y10S; Y10S930/00; Y10S930/01; Y10S930/12; Y10S930/30; Y10S930/31
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
66 words
A vector comprising the secretory signal-encoding sequence of the Bacillus licheniformis α-amylase gene; upstream from said signal-encoding sequence, a promoter sequence and a ribosome binding site sequence, transcription of said signal-encoding sequence being under the control of said promoter sequence; and downstream from said signal-encoding sequence, a site for the insertion into said vector of a heterologous DNA sequence, in reading frame with said signal-encoding sequence.
Claim 2
Dependent Claim
66 words
A vector comprising the secretory signal-encoding sequence of the Bacillus licheniformis α-amylase gene; upstream from said signal-encoding sequence, a promoter sequence and a ribosome binding site sequence, transcription of said signal-encoding sequence being under the control of said promoter sequence; and downstream from said signal-encoding sequence, a site for the insertion into said vector of a heterologous DNA sequence, in reading frame with said signal-encoding sequence.
Claim 3
Dependent Claim
38 words
The vector of claim 1, said site being located in a DNA sequence encoding a peptide or polypeptide different from the peptide or polypeptide encoded by said heterologous DNA sequence, so that said vector encodes a fusion polypeptide.
Claim 4
Dependent Claim
12 words
The vector of claim 2, said different polypeptide being B. licheniformis α-amylase.
Claim 5
Dependent Claim
18 words
The vector of claim 3, said site being at the C-terminus of the DNA sequence encoding said α-amylase.
Claim 6
Dependent Claim
34 words
The vector of claim 2, said site being located at the 3' end of said DNA sequence encoding a peptide or polypeptide different from the peptide or polypeptide encoded by said heterologous DNA sequence.
Claim 7
Dependent Claim
23 words
The vector of claim 1 wherein said promoter and ribosome binding site sequences are naturally occurring Bacillus promoter and ribosome binding site sequences.
Claim 8
Dependent Claim
12 words
A Gram positive bacterial cell transformed with the vector of claim 1.
Claim 9
Dependent Claim
13 words
The bacterial cell of claim 7, said cell being of the genus Bacillus.
Claim 10
Dependent Claim
11 words
The bacterial cell of claim 8, said cell being B. subtilis.
Claim 11
Dependent Claim
13 words
The bacterial cell of claim 7, said cell being of the genus Streptomyces.
Claim 12
Dependent Claim
39 words
The bacterial cell of claim 7 wherein said vector further comprises a DNA region homologous with a DNA region of the chromosome of said host bacterial cell, said vector being integrated into said chromosome at said region of homology.
Claim 13
Dependent Claim
11 words
A Gram-negative bacterial cell transformed with the vector of claim 1.
Claim 14
Dependent Claim
11 words
The bacterial cell of claim 12, said cell being E. coli.
Claim 15
Dependent Claim
20 words
The vector of claim 1, further comprising DNA capable of causing said vector to replicate autonomously in a Bacillus cell.
Claim 16
Dependent Claim
22 words
The vector of claim 1, further comprising DNA capable of causing said vector to integrate into the host chromosome and be amplified.
Claim 17
Dependent Claim
20 words
The vector of claim 1, further comprising DNA capable of causing said vector to replicate autonomously in a Streptomyces cell.
Claim 18
Dependent Claim
21 words
The vector of claim 1, further comprising DNA capable of causing said vector to replicate autonomously in an E. coli cell.
Claim 19
Independent Claim
145 words
A method of producing a heterologous polypeptide in a bacterial cell comprising transforming said cell with a vector comprising a DNA sequence encoding said heterologous polypeptide, said DNA sequence being positioned downstream from a DNA sequence encoding the secretory signal encoding sequence of the Bacillus licheniformis α-amylase gene, said DNA encoding said secretory signal being positioned downstream from a promoter sequence and a ribosome binding site sequence capable of functioning in said bacterial cell, transcription of said DNA sequence encoding said signal sequence and said heterologous polypeptide-encoding sequence being under the control of said promoter sequence, said secretory signal encoding sequence and said heterologous polypeptide-encoding sequence together encoding a polypeptide having a signal sequence capable of effecting the secretion from said cell of said heterologous polypeptide, culturing said cell to produce and secrete said heterologous polypeptide from said cell, and recovering said secreted heterologous polypeptide.
Claim 20
Independent Claim
145 words
A method of producing a heterologous polypeptide in a bacterial cell comprising transforming said cell with a vector comprising a DNA sequence encoding said heterologous polypeptide, said DNA sequence being positioned downstream from a DNA sequence encoding the secretory signal encoding sequence of the Bacillus licheniformis α-amylase gene, said DNA encoding said secretory signal being positioned downstream from a promoter sequence and a ribosome binding site sequence capable of functioning in said bacterial cell, transcription of said DNA sequence encoding said signal sequence and said heterologous polypeptide-encoding sequence being under the control of said promoter sequence, said secretory signal encoding sequence and said heterologous polypeptide-encoding sequence together encoding a polypeptide having a signal sequence capable of effecting the secretion from said cell of said heterologous polypeptide, culturing said cell to produce and secrete said heterologous polypeptide from said cell, and recovering said secreted heterologous polypeptide.
Claim 21
Dependent Claim
99 words
A Streptomyces cell transformed with a vector comprising a DNA sequence encoding the secretory signal-encoding sequence of the Bacillus licheniformis α-amylase gene; upstream from said signal-encoding sequence, a promoter sequence and a ribosome binding site sequence, transcription of said signal-encoding sequence being under the control of said promoter sequence; and downstream from said signal-encoding sequence, a site for the insertion into said vector of a heterologous DNA sequence, in reading frame with said signal-encoding sequence, said heterologous DNA sequence being inserted at said site, and transcription of said heterologous DNA sequence being under the control of said promoter sequence.
Claim 22
Dependent Claim
99 words
A Streptomyces cell transformed with a vector comprising a DNA sequence encoding the secretory signal-encoding sequence of the Bacillus licheniformis α-amylase gene; upstream from said signal-encoding sequence, a promoter sequence and a ribosome binding site sequence, transcription of said signal-encoding sequence being under the control of said promoter sequence; and downstream from said signal-encoding sequence, a site for the insertion into said vector of a heterologous DNA sequence, in reading frame with said signal-encoding sequence, said heterologous DNA sequence being inserted at said site, and transcription of said heterologous DNA sequence being under the control of said promoter sequence.
References Cited (Backward Citations)
EP0021571A1
Cited
EP0032238A2
Cited
GB2069503A
Cited
WO1982000341A1
Cited
GB2133408A
Cited
GB2091268A
Cited
EP0057976A2
Cited
EP0063494A2
Cited
GB2121051A
Cited
US4559300A
Cited
EP0114695A2
Cited
EP0127328A2
Cited
EP0133756A2
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-09-06 | 1992-03-07 | 1992-09-06 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-09-06 | 1996-03-07 | 1996-09-06 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-09-06 | 2000-03-07 | 2000-09-06 | $7700.00 | Window Closed / Lapsed |