US4758512A
Expired / Lapsed
US
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Hosts and methods for producing recombinant products in high yields
Assignee: Biogen MA Inc
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Filing: 1984-03-06
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Granted: 1988-07-19
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Expires: 2004-03-06
Abstract
Abstract This invention relates to improved host organisms and methods for producing recombinant products in high yields. More particularly, the present invention relates to cell strains carrying specific mutations within their DNA sequences which cause the organism to exhibit a reduced capacity for degrading foreign products and the use of these strains to produce increased yields of genetically engineered foreign proteins, polypeptides and other products. The methods disclosed in this invention advantageously permit the production, in high yields, of foreign recombinant proteins, polypeptides or other products in hosts which do not usually produce such products.
Inventors
- 👤 Alfred L. Goldberg
- 👤 Stephen A. Goff
- 👤 Lawrence P. Casson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 23:01
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12N9/6421; C12N9/6424; C12N9/6456; C12N9/6459; A; A61; A61P; A61P7/00; A61P7/02; C07; C07K; C07K14/00; C07K14/435; C07K14/575; C07K14/65; C12N15/00; C12N15/09; C12N15/63; C12N15/67; C12N15/70; C12Y; C12Y304/00; C12Y304/21; C12Y304/21069; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/832; Y10S435/839; Y10S435/848; Y10S435/849
Claims Analysis
Total Claims: 9
Claim 1
Independent Claim
87 words
A method for producing a polypeptide in a bacterial host organism comprising the steps of: (a) inserting a non-htpR DNA sequence coding for a polypeptide into a cloning vehicle so that said DNA sequence is under the regulation of a functional expression control sequence; (b) transforming a bacterial host with said cloning vehicle that has the inserted DNA sequence therein, said host carrying a mutation in a heat-shock regulatory gene that results in a reduced proteolytic degrading capacity for said polypeptide; and (c) culturing said transformed host.
Claim 2
Independent Claim
87 words
A method for producing a polypeptide in a bacterial host organism comprising the steps of: (a) inserting a non-htpR DNA sequence coding for a polypeptide into a cloning vehicle so that said DNA sequence is under the regulation of a functional expression control sequence; (b) transforming a bacterial host with said cloning vehicle that has the inserted DNA sequence therein, said host carrying a mutation in a heat-shock regulatory gene that results in a reduced proteolytic degrading capacity for said polypeptide; and (c) culturing said transformed host.
Claim 3
Independent Claim
56 words
A method for producing a polypeptide in a bacterial host organism comprising the step of culturing a bacterial host transformed with a recombinant DNA molecule characterized by a non-htpR DNA sequence coding for said polypeptide, said host carrying a mutation in a heat-shock regulatory gene that results in a reduced proteolytic degrading capacity for said polypeptide.
Claim 4
Independent Claim
56 words
A method for producing a polypeptide in a bacterial host organism comprising the step of culturing a bacterial host transformed with a recombinant DNA molecule characterized by a non-htpR DNA sequence coding for said polypeptide, said host carrying a mutation in a heat-shock regulatory gene that results in a reduced proteolytic degrading capacity for said polypeptide.
Claim 5
Dependent Claim
25 words
The method of claim 1 or 2 wherein the bacterial host organism is selected from the group consisting of htpR mutants and htpR lon mutants.
Claim 6
Dependent Claim
36 words
The method of claim 3 wherein the htpR lon mutant host organism is selected from the group consisting of E. coli strain SG927, E. coli strain SG928, E. coli strain SG935, and E. coli strain SG936.
Claim 7
Dependent Claim
123 words
The method of claim 1 or 2 wherein said non-htpR DNA sequence is selected from the group consisting of DNA sequences that code for polypeptides displaying an immunological or biological activity of leukocyte interferon, DNA sequences that code for polypeptides displaying an immunological or biological activity of fibroblast interferon, DNA sequences that code for other interferons, DNA sequences that code for polypeptides having the antigenicity of FMDV viral antigens, DNA sequences that code for polypeptides having the antigenicity of hepatitis B viral antigens, DNA sequences that code for polypeptides displaying the biological activity of proinsulin and other human or animal hormones, and DNA sequences that code for other polypeptides and proteins displaying an immunological or biological activity of human, animal or viral products.
Claim 8
Dependent Claim
14 words
The method of claim 1 or 2 wherein the expression control sequence is heat-inducible.
Claim 9
Dependent Claim
28 words
The method of claim 1 or 2 wherein the bacterial host organism is selected from the group consisting of strains of E. coli, Bacillus subtilis, and Salmonella typhimurium.
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) | 1991-07-19 | 1992-01-18 | 1992-07-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-07-19 | 1996-01-18 | 1996-07-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-07-19 | 2000-01-18 | 2000-07-19 | $7700.00 | Window Closed / Lapsed |