Patent US4649119A successfully refreshed from Google Patents.
US4649119A
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Cloning systems for corynebacterium
Assignee: Massachusetts Institute of Technology
•
Filing: 1983-04-28
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Granted: 1987-03-10
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Expires: 2003-04-28
Abstract
Abstract The invention is a method for obtaining a chimeric plasmid which can serve as a vector for inserting one or more foreign or native genes into Corynebacterium and a second species of bacteria. The plasmid is the ligation product of plasmid DNA from Corynebacterium as well as plasmid DNA from a non-Corynebacterium species of bacteria. Also disclosed is a method for forming protoplasts from Corynebacterium organisms and transforming the protoplasts with vectors, e.g., the chimeric plasmids of the invention.
Inventors
- 👤 Anthony J. Sinskey
- 👤 Graham C. Walker
- 👤 Kanji Higashio
- 👤 Eswara A. Rao
- 👤 William G. Shanabruch
- 👤 Makoto Yoshihama
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 08:40
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/74; C12N15/77; C12N15/75; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/832; Y10S435/839; Y10S435/843
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
75 words
An essentially pure plasmid pSR1, derived from a coryneform microorganism, in an isolated form suitable for use as a cloning vector for expressing foreign and native genes in coryneform microorganisms, the plasmid having a molecular weight of about 2 megadaltons and having the following cleavage sites for restriction endonucleases: ______________________________________
Enzyme # of Cleavage Sites
______________________________________
EcoR I 0
Hind II 2
Bgl II 1
Bamh I 1
Xba I 0
Bcl I 1.
______________________________________
Claim 2
Dependent Claim
75 words
An essentially pure plasmid pSR1, derived from a coryneform microorganism, in an isolated form suitable for use as a cloning vector for expressing foreign and native genes in coryneform microorganisms, the plasmid having a molecular weight of about 2 megadaltons and having the following cleavage sites for restriction endonucleases: ______________________________________
Enzyme # of Cleavage Sites
______________________________________
EcoR I 0
Hind II 2
Bgl II 1
Bamh I 1
Xba I 0
Bcl I 1.
______________________________________
Claim 3
Dependent Claim
22 words
The plasmid according to claim 1 which is isolated from a Corynebacterium microorganism having the identifying characteristics of NRRL Deposit No. B-15340.
Claim 4
Dependent Claim
76 words
An essentially pure plasmid pBD8 derived from a Bacillus subtilus microorganism, in an isolated form suitable for use as a cloning vector for expressing foreign and native genes in Bacillus microorganisms, the plasmid having a molecular weight of about 6 megadaltons and having the following cleavage sites for restriction endonucleases: ______________________________________
Enzyme # of Cleavage Sites
______________________________________
ECoR I 1
Hind II 1
Bgl II 1
Bamh I 1
Xba I 1
Bcl II 0.
______________________________________
Claim 5
Dependent Claim
76 words
An essentially pure plasmid pBD8 derived from a Bacillus subtilus microorganism, in an isolated form suitable for use as a cloning vector for expressing foreign and native genes in Bacillus microorganisms, the plasmid having a molecular weight of about 6 megadaltons and having the following cleavage sites for restriction endonucleases: ______________________________________
Enzyme # of Cleavage Sites
______________________________________
ECoR I 1
Hind II 1
Bgl II 1
Bamh I 1
Xba I 1
Bcl II 0.
______________________________________
Claim 6
Dependent Claim
23 words
The plasmid according to claim 3 which is isolated from a Bacillus microorganism having all the identifying characteristics of NRRL Deposit No. B-15305.
Claim 7
Dependent Claim
76 words
An essentially pure plasmid pBD10 derived from a Bacillus subtilus microorganism, in an isolated form suitable for use as a cloning vector for expressing foreign and native genes in Bacillus microorganisms, the plasmid having a molecular weight of about 6 megadaltons and having the following cleavage sites for restriction endonucleases: ______________________________________
Enzyme # of Cleavage Sites
______________________________________
EcoR I 0
Hind III 0
Bgl II 1
Bamh I 1
Xba I 1
Bcl I 1.
______________________________________
Claim 8
Dependent Claim
76 words
An essentially pure plasmid pBD10 derived from a Bacillus subtilus microorganism, in an isolated form suitable for use as a cloning vector for expressing foreign and native genes in Bacillus microorganisms, the plasmid having a molecular weight of about 6 megadaltons and having the following cleavage sites for restriction endonucleases: ______________________________________
Enzyme # of Cleavage Sites
______________________________________
EcoR I 0
Hind III 0
Bgl II 1
Bamh I 1
Xba I 1
Bcl I 1.
______________________________________
Claim 9
Dependent Claim
23 words
The plasmid according to claim 5 which is isolated from a Bacillus microorganism having all the identifying characteristics of NRRL Deposit No. B-15306.
Claim 10
Dependent Claim
83 words
A chimeric plasmid pHY416 of about 5 megadaltons molecular weight prepared by isolating a first plasmid of about 2 megadaltons molecular weight from a Corynebacterium microorganism and a second plasmid of about 4 megadaltons from a Bacillus microorganism, cleaving the plasmids with a restriction endonuclease, and ligating the cleaved plasmids to one another to produce a chimeric plasmid having the following cleavage sites for restriction endonucleases: ______________________________________
Enzyme # of Cleavage Sites
______________________________________
Bcl I 2
BamH I 1
Bgl II 2.
______________________________________
Claim 11
Dependent Claim
83 words
A chimeric plasmid pHY416 of about 5 megadaltons molecular weight prepared by isolating a first plasmid of about 2 megadaltons molecular weight from a Corynebacterium microorganism and a second plasmid of about 4 megadaltons from a Bacillus microorganism, cleaving the plasmids with a restriction endonuclease, and ligating the cleaved plasmids to one another to produce a chimeric plasmid having the following cleavage sites for restriction endonucleases: ______________________________________
Enzyme # of Cleavage Sites
______________________________________
Bcl I 2
BamH I 1
Bgl II 2.
______________________________________
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1990-03-09 | 1990-09-08 | 1991-03-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1994-03-09 | 1994-09-08 | 1995-03-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1998-03-09 | 1998-09-08 | 1999-03-10 | $7700.00 | Window Closed / Lapsed |