US4683205A
Expired / Lapsed
US
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Method for transforming microorganisms
Assignee: KH Neochem Co Ltd
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Filing: 1985-02-04
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Granted: 1987-07-28
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Expires: 2005-02-04
Abstract
Abstract Disclosed is a method for transformation of microorganisms belonging to the genera Corynebacterium and Brevibacterium whereby a foreign DNA may be introduced into a host cell and autonomously replicated.
Inventors
- 👤 Ryoichi Katsumata
- 👤 Akio Ozaki
- 👤 Tetsuo Oka
- 👤 Akira Furuya
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 09:02
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/74; C12N15/77; C12N1/00; C12N1/20; C12N1/205; C12R; C12R2001/00; C12R2001/01; C12R2001/13; C12R2001/15
Claims Analysis
Total Claims: 9
Claim 1
Independent Claim
106 words
A method for transforming microorganisms, which comprises: (1) selecting a microbial strain of one of the group of genera consisting of Corynbacterium and Brevibacterium; (2) incubating protoplasts of said microbial strain and a donor deoxyribonucleic acid selected from the group consisting of chromosomal DNAs which are stabilized after cellular introduction by recombination and plasmids in the presence of polyethylene glycol or polyvinylalcohol and at least one divalent metal cation whereby to introduce said donor deoxyribonucleic acid into said protoplasts; (3) regenerating said protoplasts by culturing in a hypertonic nutrient medium; and (4) thereafter recovering a transformed strain having a phenotype derived from said donor deoxyribonucleic acid.
Claim 2
Independent Claim
106 words
A method for transforming microorganisms, which comprises: (1) selecting a microbial strain of one of the group of genera consisting of Corynbacterium and Brevibacterium; (2) incubating protoplasts of said microbial strain and a donor deoxyribonucleic acid selected from the group consisting of chromosomal DNAs which are stabilized after cellular introduction by recombination and plasmids in the presence of polyethylene glycol or polyvinylalcohol and at least one divalent metal cation whereby to introduce said donor deoxyribonucleic acid into said protoplasts; (3) regenerating said protoplasts by culturing in a hypertonic nutrient medium; and (4) thereafter recovering a transformed strain having a phenotype derived from said donor deoxyribonucleic acid.
Claim 3
Dependent Claim
24 words
A method according to claim 1 wherein said divalent metal cation is selected from the group consisting of calcium, magnesium, manganese, barium and strontium.
Claim 4
Dependent Claim
23 words
A method according to claim 2 wherein said divalent metal cation is present at a concentration of from about 1 to 100 mM.
Claim 5
Dependent Claim
19 words
A method according to claim 1 wherein said polyethyleneglycol is present at a concentration of from 5 to 60%.
Claim 6
Dependent Claim
19 words
A method according to claim 1 wherein said polyvinylalcohol is present at a concentration of from 1 to 20%.
Claim 7
Dependent Claim
17 words
A method according to claim 1 wherein said donor deoxyribonucleic acid is a plasmid or a phage.
Claim 8
Dependent Claim
21 words
A method according to claim 6 wherein said plasmid is derived from a microorganism belonging to the genus Corynebacterium or Brevibacterium.
Claim 9
Dependent Claim
12 words
A method according to claim 7 wherein said plasmid is plasmid pCG4.
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) | 1990-07-27 | 1991-01-26 | 1991-07-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1994-07-27 | 1995-01-26 | 1995-07-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1998-07-27 | 1999-01-26 | 1999-07-28 | $7700.00 | Window Closed / Lapsed |