US4348478A
Expired / Lapsed
US
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Method for preparation of a recombinant DNA phage
Assignee: Noda Institute for Scientific Research
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Filing: 1979-10-22
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Granted: 1982-09-07
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Expires: 1999-10-22
Abstract
Abstract A method for the preparation of a novel recombinant DNA, which comprises (1) cleaving with an endonuclease a temperate phage DNA having an endonuclease-sensitive region not in the DNA segment participating in the replication of phage DNA and the integration of phage DNA into a host chromosome but at least in the DNA segment carrying genetic information for the coat protein production and another DNA carrying intended genetic information, (2) adding DNA-ligase to the mixture of both cleft DNA's, and (3) recovering from the mixture a phage DNA having its coat protein producing ability deleted by the replacement of the DNA segments carrying genetic information for the coat protein production with a DNA fragment carrying the intended information.
Inventors
- 👤 Eiichi Nakano
- 👤 Tsutomu Masuda
- 👤 Narimasa Saito
- 👤 Danji Fukushima
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-28 13:38
CPC Classifications
C; C12; C12N; C12N15/00
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
118 words
A method for the preparation of a novel recombinant DNA, which comprises (1) cleaving with an endonuclease a temperate phage DNA having an endonuclease-sensitive region not in the DNA segment participating in the replication of phage DNA and the integration of phage DNA into a host chromosome but at least in the DNA segment carrying genetic information for the coat protein production and another DNA carrying intended genetic information, (2) adding DNA-ligase to the mixture of both cleft DNA's, and (3) recovering from the mixture a phage DNA having its coat protein producing ability deleted by the replacement of the DNA segment carrying genetic information for the coat protein production with a DNA fragment carrying the intended information.
Claim 2
Independent Claim
118 words
A method for the preparation of a novel recombinant DNA, which comprises (1) cleaving with an endonuclease a temperate phage DNA having an endonuclease-sensitive region not in the DNA segment participating in the replication of phage DNA and the integration of phage DNA into a host chromosome but at least in the DNA segment carrying genetic information for the coat protein production and another DNA carrying intended genetic information, (2) adding DNA-ligase to the mixture of both cleft DNA's, and (3) recovering from the mixture a phage DNA having its coat protein producing ability deleted by the replacement of the DNA segment carrying genetic information for the coat protein production with a DNA fragment carrying the intended information.
Claim 3
Dependent Claim
14 words
A method according to claim 1, wherein the temperate phage is a lambdoid phage.
Claim 4
Dependent Claim
35 words
A method according to claim 2, wherein the lambdoid phage is lamda (IFO 20016) 434(IFO 20018), 82(IFO 20019), φ80(IFO 20020), φ170(IFO 20021), E. coli K12 strain W3110(φ80)(ATCC 31277) or E. coli K12 strain W3350(λcI857)(ATCC 31278).
Claim 5
Dependent Claim
78 words
A method according to claim 1, wherein the phage DNA having an endonuclease-sensitive region not in the DNA segment participating in temperate phage DNA replication and integration of DNA into a host chromosome but at least in the DNA segment carrying genetic information for the coat protein production is a DNA obtained by mating an endonuclease-resistant temperate phage with a temperate phage having an endonuclease-sensitive region in the DNA segment participating in the production of a coat protein.
Claim 6
Dependent Claim
23 words
A method according to claim 4, wherein the endonuclease-resistant temperate phage is a phage obtained from a deletion mutant by microbial concentration technique.
Claim 7
Dependent Claim
70 words
A method according to claim 1, wherein the phage DNA having an endonuclease-sensitive region not in the DNA segment participating in phage DNA replication and integration of DNA into a host chromosome but at least in the DNA segment carrying genetic information for the coat protein production is a DNA obtained by mating a phage having an endonuclease-sensitive region only in the middle of phage DNA with a transducing phage.
Claim 8
Dependent Claim
33 words
A method according to claim 6, wherein the phage having an endonuclease-sensitive region only in the middle of phage DNA is one obtained by mating an endonuclease-resistant phage with the parent strain thereof.
Claim 9
Dependent Claim
24 words
A method according to claim 1, wherein the DNA carrying intended genetic information is a DNA originated from molds, higher organisms or transducing phages.
Claim 10
Dependent Claim
13 words
A method according to claim 1, wherein the endonuclease is a restriction enzyme.
Claim 11
Dependent Claim
14 words
A method according to claim 9, the restriction enzyme is EcoRI, BamI or HindIII.
Claim 12
Dependent Claim
25 words
A method according to claim 1, wherein the DNA ligase concentration is 1 to 10 units/ml to the DNA concentration of 10 to 80 μg/ml.
Claim 13
Dependent Claim
21 words
A method according to claim 1, wherein the DNA ligase is E. coli DNA ligase or T 4 phage DNA ligase.
Claim 14
Dependent Claim
21 words
A method according to claim 11, wherein the mixture is kept at 0° to 10° C. for 1 to 14 days.
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) | 1985-09-06 | 1986-03-08 | 1986-09-07 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1989-09-06 | 1990-03-08 | 1990-09-07 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1993-09-06 | 1994-03-08 | 1994-09-07 | $7700.00 | Window Closed / Lapsed |