US5028530A
Expired / Lapsed
US
⏱️ 0.0 years remaining
AraB promoters and method of producing polypeptides, including cecropins, by microbiological techniques
Assignee: Xoma Technology Ltd
•
Filing: 1990-02-05
•
Granted: 1991-07-02
•
Expires: 2010-02-05
Abstract
Abstract A polynucleotide molecule expressible in a given host comprising the sequence of the araB promoter operably linked to a gene which is heterologous to said host. The heterologous gene codes for a peptide that is biologically active. The invention also relates to a genetic construct which comprises a first genetic sequence coding for cecropin operably linked to a second genetic sequence coding for a polypeptide which is capable of suppressing the bactericidal effect of the resulting fusion protein towards an otherwise cecropin sensitive bacterium.
Inventors
- 👤 Jiunu Lai
- 👤 Jar-How Lee
- 👤 Yun-Long Lin
- 👤 Dan Ray
- 👤 Gary Wilcox
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 09:30
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/43504; C07K14/43563; C07K14/575; C07K14/585; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/62; C12N15/63; C12N15/70; C07K2319/00
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
66 words
A replicable expression vehicle comprising: (a) the sequence of an araB promoter from a member of the Enterobacteriaceae family; and, (b) the coding sequence of a gene, wherein said gene is operably linked to said promoter such that said gene is expressible in a given araC + host for said vehicle by induction of said araB promoter and wherein said gene is heterologous to said host.
Claim 2
Dependent Claim
66 words
A replicable expression vehicle comprising: (a) the sequence of an araB promoter from a member of the Enterobacteriaceae family; and, (b) the coding sequence of a gene, wherein said gene is operably linked to said promoter such that said gene is expressible in a given araC + host for said vehicle by induction of said araB promoter and wherein said gene is heterologous to said host.
Claim 3
Dependent Claim
25 words
The vehicle of claim 41 wherein said gene codes for a polypeptide selected from the group consisting of enzymes, hormones, antibodies, hemoglobin, and structural proteins.
Claim 4
Dependent Claim
12 words
The vehicle of claim 41 wherein said gene codes for a cecropin.
Claim 5
Dependent Claim
10 words
The vehicle of claim 41 which is a DNA molecule.
Claim 6
Dependent Claim
11 words
The vehicle of claim 41 which is a double-stranded DNA molecule.
Claim 7
Dependent Claim
13 words
The vehicle of claim 1, wherein said araB promoter is from S. typhimurium.
Claim 8
Dependent Claim
14 words
The vehicle of claim 1, wherein said araB promoter is heterologous to said host.
Claim 9
Dependent Claim
9 words
The vehicle of claim 42 which is a plasmid.
Claim 10
Dependent Claim
9 words
The vehicle of claim 42 which is a bacteriophage.
Claim 11
Dependent Claim
12 words
The plasmid of claim 7 which also contains a phenotypic selection marker.
Claim 12
Dependent Claim
11 words
The plasmid of claim 9 wherein said marker is antibiotic resistance.
Claim 13
Dependent Claim
9 words
A bacterium transformed with the vehicle of claim 41.
Claim 14
Dependent Claim
121 words
A process for producing a biologically active heterologous peptide in a transformed araC + bacterium, wherein said process comprises: (a) inserting a polynucleotide molecule into a replicable expression vehicle, wherein said polynucleotide molecule comprises an araB promoter from a member of the Enterobacteriaceae family, and wherein said araB promoter is operably linked to a gene heterologous to said araC + bacterium, said gene encoding said peptide; (b) transforming said araC + bacterium with said expression vehicle containing said polynucleotide molecule of part (a); (c) culturing said araC + bacterium of part (b) under culturing conditions wherein said conditions induce expression of said gene encoding said peptide which is operably-linked to said araB promoter; and (d) recovering said peptide from the culture.
Claim 15
Dependent Claim
121 words
A process for producing a biologically active heterologous peptide in a transformed araC + bacterium, wherein said process comprises: (a) inserting a polynucleotide molecule into a replicable expression vehicle, wherein said polynucleotide molecule comprises an araB promoter from a member of the Enterobacteriaceae family, and wherein said araB promoter is operably linked to a gene heterologous to said araC + bacterium, said gene encoding said peptide; (b) transforming said araC + bacterium with said expression vehicle containing said polynucleotide molecule of part (a); (c) culturing said araC + bacterium of part (b) under culturing conditions wherein said conditions induce expression of said gene encoding said peptide which is operably-linked to said araB promoter; and (d) recovering said peptide from the culture.
Claim 16
Dependent Claim
156 words
A process for producing a heterologous peptide, wherein said process comprises the steps of: (a) incubating a transformed araC + bacterium in a culture medium under selected growth conditions until late exponential growth phase is reached, wherein said bacterium has been transformed with a polynucleotide molecule comprising an araB promoter from a member of the Enterobacteriaceae family operably linked to a gene heterologous to said bacterium and coding for a peptide; (b) transferring said bacterium of step (a) to fresh culture medium and incubating said bacterium for a period of time sufficient to allow further growth; (c) inoculating said culture medium of step (b) with production medium and incubating said bacterium in said production medium at fermentation conditions to achieve maximal bacterial growth and for a time sufficient to achieve a selected cell density; (d) adding an amount of L-arabinose effective to produce substantial levels of said peptide in said medium, and (e) recovering said peptide.
Claim 17
Dependent Claim
156 words
A process for producing a heterologous peptide, wherein said process comprises the steps of: (a) incubating a transformed araC + bacterium in a culture medium under selected growth conditions until late exponential growth phase is reached, wherein said bacterium has been transformed with a polynucleotide molecule comprising an araB promoter from a member of the Enterobacteriaceae family operably linked to a gene heterologous to said bacterium and coding for a peptide; (b) transferring said bacterium of step (a) to fresh culture medium and incubating said bacterium for a period of time sufficient to allow further growth; (c) inoculating said culture medium of step (b) with production medium and incubating said bacterium in said production medium at fermentation conditions to achieve maximal bacterial growth and for a time sufficient to achieve a selected cell density; (d) adding an amount of L-arabinose effective to produce substantial levels of said peptide in said medium, and (e) recovering said peptide.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1994-07-01 | 1994-12-31 | 1995-07-02 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-07-01 | 1998-12-31 | 1999-07-02 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-07-01 | 2002-12-31 | 2003-07-02 | $7700.00 | Window Closed / Lapsed |