US4499188A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Bacterial production of heterologous polypeptides under the control of a repressible promoter-operator
Assignee: Novartis Vaccines and Diagnostics Inc
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Filing: 1982-05-05
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Granted: 1985-02-12
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Expires: 2002-05-05
Abstract
Abstract A process for bacterially producing heterologous polypeptides, particularly those such as human IFN-β that inhibit bacterial growth, in which bacteria that have been transformed to express the heterologous polypeptide under the control of a trp promoter-operator are cultivated in a known volume of medium containing an excess of a preferred carbon source such as glucose and a predetermined amount of tryptophan that corresponds approximately to the amount of tryptophan contained in the bacteria in the volume of medium at a predetermined elevated cellular density, whereby expression of the heterologous polypeptide is substantially repressed until the bacteria grow to approximately the predetermined elevated cellular density and is thereafter automatically derepressed to permit expression of the heterologous polypeptide.
Inventors
- 👤 Michael W. Konrad
- 👤 David F. Mark
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-27 09:27
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/555; C07K14/565; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/70; C12N15/71; Y; Y10; Y10S; Y10S930/00; Y10S930/01; Y10S930/14; Y10S930/142
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
147 words
A process for bacterially producing a heterologous polypeptide comprising cultivating bacteria transformed to express said heterologous polypeptide under the control of a bacterial promoteroperator that (i) normally controls the expression of the structural genes that encode the polypeptides of a biosynthetic pathway that produces a product that is used by the bacteria in growth, (ii) is repressed by said product through activation of a repressor molecule by said product, and (iii) provides efficient expression of the structural genes under its control when derepressed in a volume of a culture medium containing an excess of essential nutrients and a predetermined amount of said product that corresponds approximately to the amount of said product contained in the bacteria in said volume at a predetermined, elevated cellular density for a time sufficient to permit growth of the bacteria to the predetermined cellular density and expression of the heterologous polypeptide thereafter.
Claim 2
Dependent Claim
147 words
A process for bacterially producing a heterologous polypeptide comprising cultivating bacteria transformed to express said heterologous polypeptide under the control of a bacterial promoteroperator that (i) normally controls the expression of the structural genes that encode the polypeptides of a biosynthetic pathway that produces a product that is used by the bacteria in growth, (ii) is repressed by said product through activation of a repressor molecule by said product, and (iii) provides efficient expression of the structural genes under its control when derepressed in a volume of a culture medium containing an excess of essential nutrients and a predetermined amount of said product that corresponds approximately to the amount of said product contained in the bacteria in said volume at a predetermined, elevated cellular density for a time sufficient to permit growth of the bacteria to the predetermined cellular density and expression of the heterologous polypeptide thereafter.
Claim 3
Dependent Claim
17 words
The process of claim 1 wherein the promoter-operator is the trp promoter-operator and the product is tryptophan.
Claim 4
Dependent Claim
17 words
The process of claim 1 or 2 wherein the heterologous polypeptide affects bacterial growth or viability adversely.
Claim 5
Dependent Claim
14 words
The process of claim 1 or 2 wherein the heterologous polypeptide is human IFN-β.
Claim 6
Dependent Claim
27 words
The process of claim 1 or 2 wherein the heterologous polypeptide is human IFN-β, the bacteria are E.coli, and the trp promoter-operator is the E.coli trp promoter-operator.
Claim 7
Dependent Claim
22 words
The process of claim 5 wherein the predetermined amount of tryptophan is defined by the formula: mg tryptophan/l of medium ≈5.3×predetermined turbidity.
Claim 8
Dependent Claim
18 words
The process of claim 1 or 2 wherein the predetermined elevated cellular density is substantially maximum cellular density.
Claim 9
Dependent Claim
29 words
The process of claim 2 wherein the predetermined elevated cellular density as measured by the turbidity of the medium is in the range of about 10 to about 100.
Claim 10
Dependent Claim
33 words
The process of claim 8 wherein the bacteria are E.coli and the predetermined amount of tryptophan is in the corresponding range of about 53 mg/l of medium to about 530 mg/l of medium.
Claim 11
Dependent Claim
15 words
The process of claim 9 wherein the bacteria have been transformed with the plasmid pβl-trp.
References Cited (Backward Citations)
EP0036776A2
Cited
US4374927A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1988-02-12 | 1988-08-13 | 1989-02-12 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-02-12 | 1992-08-13 | 1993-02-12 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-02-12 | 1996-08-13 | 1997-02-12 | $7700.00 | Window Closed / Lapsed |