US4460688A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Plasmid and its use
Assignee: Gist Brocades NV
•
Filing: 1981-12-21
•
Granted: 1984-07-17
•
Expires: 2001-12-21
Abstract
Abstract Plasmid conferring resistance to streptomycin (SmR) and neomycin (NeoR) upon its host (e.g. a Bacillus, Staphylococcus or Escherichia coli) and which after having taken up a foreign DNA fragment at one of its restriction sites with loss of its SmR or NeoR phenotype, is still capable of replicating and expressing genetic information in its host.
Inventors
- 👤 Johan P. M. Sanders
- 👤 Andrew J. P. Docherty
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 20:08
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/74; C12N15/75
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
143 words
A process for preparing a plasmid which confers resistance to streptomycin (Sm R ) and neomycin (Neo R ) upon a Bacillus host and which after having taken up a foreign DNA fragment at one of its restriction sites with loss of its Sm R or Neo R phenotype but not both, is still capable of replicating and expressing genetic information in its host comprising preparing plasmid pUB 109 conferring resistance to streptomycin, (Sm R ), preparing plasmid pUB 110 conferring resistance to neomycin, (Neo R ), introducing each of said plasmids into a Bacillus host, isolating said plasmids from said host, restricting said plasmids with restriction enzyme Xba I not interfering with the gene properties as to Sm R and Neo R of the starting plasmids recombining the restricted plasmids and isolating the recombined plasmid conferring Sm R and Neo R .
Claim 2
Dependent Claim
143 words
A process for preparing a plasmid which confers resistance to streptomycin (Sm R ) and neomycin (Neo R ) upon a Bacillus host and which after having taken up a foreign DNA fragment at one of its restriction sites with loss of its Sm R or Neo R phenotype but not both, is still capable of replicating and expressing genetic information in its host comprising preparing plasmid pUB 109 conferring resistance to streptomycin, (Sm R ), preparing plasmid pUB 110 conferring resistance to neomycin, (Neo R ), introducing each of said plasmids into a Bacillus host, isolating said plasmids from said host, restricting said plasmids with restriction enzyme Xba I not interfering with the gene properties as to Sm R and Neo R of the starting plasmids recombining the restricted plasmids and isolating the recombined plasmid conferring Sm R and Neo R .
Claim 3
Dependent Claim
13 words
Process according to claim 1 wherein the Bacillus is B.subtilis, B.licheniformis or B.amyloliquefaciens.
Claim 4
Dependent Claim
14 words
Process according to claim 2 wherein the Bacillus is the restrictionless B.subtilis 1G 20.
Claim 5
Dependent Claim
13 words
Process according to claim 1 in which the product is pUB1654 (CBS 237.79).
Claim 6
Dependent Claim
40 words
The process according to claim 1 comprising the additional step of inserting a foreign DNA fragment at one of the resulting unique restriction sites of the recombined plasmid located within either the Sm R gene or the Neo R gene.
Claim 7
Dependent Claim
41 words
The process of claim 2 or 3 comprising the additional step of inserting a foreign DNA fragment at one of the resulting unique restriction sites of the recombined plasmid located within either the Sm R gene or the Neo R gene.
Claim 8
Dependent Claim
14 words
Process according to claim 5 wherein the restriction site is the Bql II site.
Claim 9
Dependent Claim
17 words
Process according to claim 1 comprising the additional step of introducing the recombined plasmid into a host.
Claim 10
Dependent Claim
14 words
Process according to claim 8 wherein the host is a B.subtilis, B.licheniformis or B.amyloliquefaciens.
Claim 11
Dependent Claim
47 words
Process for replicating and expressing a foreign DNA fragment in a plasmid vehicle in a host according to the process defined in any one of claim 5 to 8 and 9 comprising incubating said host under conditions suitable for replication and expression of said foreign DNA fragments.
Claim 12
Dependent Claim
14 words
Process according to claim 5 wherein the restriction site is the Hind III site.
Claim 13
Dependent Claim
23 words
The process of claim 2, 3, 5, 6, 7 or 11 comprising the additional step of introducing the recombined plasmid into a host.
Claim 14
Dependent Claim
14 words
Process according to claim 5 wherein the restriction site is the Eco RI site.
References Cited (Backward Citations)
US4237224A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-07-17 | 1988-01-16 | 1988-07-17 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-07-17 | 1992-01-16 | 1992-07-17 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-07-17 | 1996-01-16 | 1996-07-17 | $7700.00 | Window Closed / Lapsed |