US4617266A
Expired / Lapsed
US
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Production of Protein A
Assignee: Genex Corp
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Filing: 1983-04-28
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Granted: 1986-10-14
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Expires: 2003-04-28
Abstract
Abstract Novel Protein A-producing Gram-positive bacterial strains and methods for their preparation are disclosed. Also disclosed are methods for producing Protein A using the novel Gram-positive strains.
Inventors
- 👤 Stephen R. Fahnestock
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 01:40
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/195; C07K14/305; C07K14/31; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/74; C12N15/75; C12N15/87; C12N15/90; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/832; Y10S435/839
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
89 words
A plasmid vector comprising a DNA sequence specifying Protein A; expression signals operably linked to said DNA sequence for directing expression of the Protein A DNA sequence in a Gram-positive microorganism of the species "B. subtilis"; a selectable phenotypic marker which is expressable in a Gram-positive microorganism of the species "B. subtilis"; and a region of sequence homology with a chromosome of a Gram-positive microorganism of the species "B. substilis", said region of sequence homology being capable of permitting integration of the Protein A DNA sequence into said chromosome.
Claim 2
Dependent Claim
89 words
A plasmid vector comprising a DNA sequence specifying Protein A; expression signals operably linked to said DNA sequence for directing expression of the Protein A DNA sequence in a Gram-positive microorganism of the species "B. subtilis"; a selectable phenotypic marker which is expressable in a Gram-positive microorganism of the species "B. subtilis"; and a region of sequence homology with a chromosome of a Gram-positive microorganism of the species "B. substilis", said region of sequence homology being capable of permitting integration of the Protein A DNA sequence into said chromosome.
Claim 3
Dependent Claim
24 words
The plasmid vector of claim 1 further comprising a selectable phenotypic marker which is expressable in E. coli and a functional E. coli replicon.
Claim 4
Independent Claim
91 words
A method of producing Protein A comprising cultivating in a nutrient medium a transformed Gram-positive microorganism of the species "B. subtilis" transformed by a vector containing a nucleotide sequence coding for Protein A and expression signals operably linked to said DNA sequence for directing expression of Protein A in the transformed microorganism, to produce protein A; wherein the vector contains a DNA fragment homologous to a region of the chromosome of said microorganism, and the nucleotide sequence coding for Protein A is integrated into the microorganism host chromosome under recombination conditions.
Claim 5
Independent Claim
91 words
A method of producing Protein A comprising cultivating in a nutrient medium a transformed Gram-positive microorganism of the species "B. subtilis" transformed by a vector containing a nucleotide sequence coding for Protein A and expression signals operably linked to said DNA sequence for directing expression of Protein A in the transformed microorganism, to produce protein A; wherein the vector contains a DNA fragment homologous to a region of the chromosome of said microorganism, and the nucleotide sequence coding for Protein A is integrated into the microorganism host chromosome under recombination conditions.
Claim 6
Dependent Claim
17 words
The method of claim 3 wherein the vector is a plasmid capable of replication in E. coli.
Claim 7
Dependent Claim
50 words
The method of claim 4 wherein plasmids are prepared which contain DNA fragments homologous to different regions of the chromosome of the Gram-positive microorganism, and such plasmids are linearly integrated into their respective homologous regions of the host chromosome under recombination conditions resulting in multiple insertions into the host chromosome.
Claim 8
Dependent Claim
26 words
The method of claim 4 wherein the plasmid contains a DNA fragment homologous to sequences of the chromosomes of more than one species of Gram-positive bacteria.
Claim 9
Dependent Claim
21 words
The method of claim 4 wherein the plasmid contains more than one copy of the nucleotide sequence coding for Protein A.
Claim 10
Dependent Claim
21 words
The method of claim 4 wherein the plasmid contains more than one copy of the nucleotide sequence coding for Protein A.
Claim 11
Dependent Claim
20 words
The method of claim 4 wherein the plasmid contains two tandem copies of the nucleotide sequence coding for Protein A.
Claim 12
Dependent Claim
41 words
A protein A-producing strain of a Gram-positive microorganism of the species "B. subtilis", wherein the chromosome of said microorganism comprises a heterologous Protein A gene under the requesting central of expression signals capable of directing expression of the Protein A gene.
Claim 13
Dependent Claim
41 words
A protein A-producing strain of a Gram-positive microorganism of the species "B. subtilis", wherein the chromosome of said microorganism comprises a heterologous Protein A gene under the requesting central of expression signals capable of directing expression of the Protein A gene.
Claim 14
Independent Claim
27 words
A method of producing Protein A comprising cultivating in a nutrient medium a strain of Bacillus subtilis, having the identifying characteristics of strain GX3305 (ATCC No. 39345).
Claim 15
Independent Claim
27 words
A method of producing Protein A comprising cultivating in a nutrient medium a strain of Bacillus subtilis, having the identifying characteristics of strain GX3305 (ATCC No. 39345).
Claim 16
Dependent Claim
7 words
Bacillus subtilis strain GX3305 (ATCC No. 39345).
Claim 17
Dependent Claim
7 words
Bacillus subtilis strain GX3305 (ATCC No. 39345).
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1989-10-13 | 1990-04-14 | 1990-10-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-10-13 | 1994-04-14 | 1994-10-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-10-13 | 1998-04-14 | 1998-10-14 | $7700.00 | Window Closed / Lapsed |