US4259444A
Expired / Lapsed
US
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Microorganisms having multiple compatible degradative energy-generating plasmids and preparation thereof
Assignee: General Electric Co
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Filing: 1972-06-07
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Granted: 1981-03-31
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Expires: 1992-06-07
Abstract
Abstract Unique microorganisms have been developed by the application of genetic engineering techniques. These microorganisms contain at least two stable (compatible) energy-generating plasmids, these plasmids specifying separate degradative pathways. The techniques for preparing such multi-plasmid strains from bacteria of the genus Pseudomonas are described. Living cultures of two strains of Pseudomonas (P. aeruginosa [NRRL B-5472] and P. putida [NRRL B-5473]) have been deposited with the United States Department of Agriculture, Agricultural Research Service, Northern Marketing and Nutrient Research Division, Peoria, Ill. The P. aeruginosa NRRL B-5472 was derived from Pseudomonas aeruginosa strain 1c by the genetic transfer thereto, and containment therein, of camphor, octane, salicylate and naphthalene degradative pathways in the form of plasmids. The P. putida NRRL B-5473 was derived from Pseudomonas putida strain PpG1 by genetic transfer thereto, and containment therein, of camphor, salicylate and naphthalene degradative pathways and drug resistance factor RP-1, all in the form of plasmids.
Inventors
- 👤 Ananda M. Chakrabarty
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-28 11:01
CPC Classifications
C; C12; C12N; C12N15/00; Y; Y10; Y10S; Y10S423/00; Y10S423/09; Y10S423/17; Y10S435/00; Y10S435/82; Y10S435/8215; Y10S435/822; Y10S435/874; Y10S435/875; Y10S435/877
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
24 words
A bacterium from the genus Pseudomonas containing therein at least two stable energy-generating plasmids, each of said plasmids providing a separate hydrocarbon degradative pathway.
Claim 2
Dependent Claim
24 words
A bacterium from the genus Pseudomonas containing therein at least two stable energy-generating plasmids, each of said plasmids providing a separate hydrocarbon degradative pathway.
Claim 3
Dependent Claim
26 words
The Pseudomonas bacterium of claim 1 wherein the hydrocarbon degradative pathways are selected from the group consisting of linear aliphatic, cyclic aliphatic, aromatic and polynuclear aromatic.
Claim 4
Dependent Claim
14 words
The Pseudomonas bacterium of claim 1, said bacterium being of the specie P. aeruginosa.
Claim 5
Dependent Claim
17 words
The P. aeruginosa bacterium of claim 3 wherein the bacterium contains CAM, OCT, SAL and NPL plasmids.
Claim 6
Dependent Claim
14 words
The Pseudomonas bacterium of claim 1, said bacterium being of the specie P. putida.
Claim 7
Dependent Claim
17 words
The P. putida bacterium of claim 5 wherein the bacterium contains CAM, SAL, NPL and RP-1 plasmids.
Claim 8
Dependent Claim
48 words
An inoculum for the degradation of a preselected substrate comprising a complex or mixture of hydrocarbons, said inoculum consisting essentially of bacteria of the genus Pseudomonas at least some of which contain at least two stable energy-generating plasmids, each of said plasmids providing a separate hydrocarbon degradative pathway.
Claim 9
Dependent Claim
48 words
An inoculum for the degradation of a preselected substrate comprising a complex or mixture of hydrocarbons, said inoculum consisting essentially of bacteria of the genus Pseudomonas at least some of which contain at least two stable energy-generating plasmids, each of said plasmids providing a separate hydrocarbon degradative pathway.
Claim 10
Dependent Claim
25 words
The inoculum of claim 7 wherein the hydrocarbon degradative pathways are selected from the group consisting of linear aliphatic, cyclic aliphatic, aromatic and polynuclear aromatic.
Claim 11
Dependent Claim
18 words
The inoculum of claim 8 wherein the bacteria having multiple energy-generating plasmids are of the specie P. aeruginosa.
Claim 12
Dependent Claim
18 words
The inoculum of claim 8 wherein the bacteria having multiple energy-generating plasmids are of the specie P. putida.
Claim 13
Dependent Claim
119 words
In the process in which a first energy-generating plasmid specifying a degradative pathway is transferred by conjugation from a donor Pseudomonas bacterium to a recipient Pseudomonas bacterium containing at least one energy-generating plasmid that is incompatible with said first plasmid, said transfer occurring in the quiescent state after the mixing of substantially equal volumes of cultures of said donor and said recipient, each culture presenting the respective organisms in a complex nutrient liquid medium at a population density of at least about 1,000,000 cells/ml, the improvement wherein after conjugation has occurred, the multi-plasmid conjugatant bacteria are subjected to DNA-cleaving radiation in a dosage sufficient to fuse the first plasmid and the plasmid incompatible therewith located in the same cell.
Claim 14
Dependent Claim
119 words
In the process in which a first energy-generating plasmid specifying a degradative pathway is transferred by conjugation from a donor Pseudomonas bacterium to a recipient Pseudomonas bacterium containing at least one energy-generating plasmid that is incompatible with said first plasmid, said transfer occurring in the quiescent state after the mixing of substantially equal volumes of cultures of said donor and said recipient, each culture presenting the respective organisms in a complex nutrient liquid medium at a population density of at least about 1,000,000 cells/ml, the improvement wherein after conjugation has occurred, the multi-plasmid conjugatant bacteria are subjected to DNA-cleaving radiation in a dosage sufficient to fuse the first plasmid and the plasmid incompatible therewith located in the same cell.
Claim 15
Dependent Claim
12 words
The improvement of claim 11 wherein the DNA-cleaving radiation is UV radiation.
Claim 16
Dependent Claim
25 words
The improvement of claim 12 wherein the first plasmid provides the degradative pathway for camphor and the recipient Pseudomonas contains the degradative pathway for n-octane.
Claim 17
Dependent Claim
69 words
An inoculated medium for the degradation of liquid hydrocarbon substrate material floating on water, said inoculated medium comprising a carrier material able to float on water and bacteria from the genus Pseudomonas carried thereby, at least some of said bacteria each containing at least two stable energy-generating plasmids, each of said plasmids providing a separate hydrocarbon degradative pathway and said carrier material being able to absorb said hydrocarbon material.
Claim 18
Dependent Claim
69 words
An inoculated medium for the degradation of liquid hydrocarbon substrate material floating on water, said inoculated medium comprising a carrier material able to float on water and bacteria from the genus Pseudomonas carried thereby, at least some of said bacteria each containing at least two stable energy-generating plasmids, each of said plasmids providing a separate hydrocarbon degradative pathway and said carrier material being able to absorb said hydrocarbon material.
Claim 19
Dependent Claim
12 words
The inoculated medium of claim 14 wherein the carrier material is straw.
Claim 20
Dependent Claim
26 words
The inoculated medium of claim 14 wherein the hydrocarbon degradative pathways are selected from the group consisting of linear aliphatic, cyclic aliphatic, aromatic and polynuclear aromatic.
Claim 21
Dependent Claim
15 words
The inoculated medium of claim 14 wherein the bacteria are of the specie P. aeruginosa.
Claim 22
Dependent Claim
15 words
The inoculated medium of claim 14 wherein the bacteria are of the specie P. putida.
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) | 1984-03-30 | 1984-09-29 | 1985-03-31 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1988-03-30 | 1988-09-29 | 1989-03-31 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1992-03-30 | 1992-09-29 | 1993-03-31 | $7700.00 | Window Closed / Lapsed |