US4599308A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Protein from SV40 recombinants
Assignee: Individual
•
Filing: 1983-09-29
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Granted: 1986-07-08
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Expires: 2003-09-29
Abstract
Abstract An E. Coli plasmid SV40 vector recombinant is cloned to a gene of interest and amplified in bacteria. The SV40 vector-gene of interest can be introduced into eukaryotic cells by transformation or transfection and the gene of interest produces its protein product.
Inventors
- 👤 Dean H. Hamer
- 👤 Marian Kaehler
- 👤 Philip Leder
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 14:21
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85; C12N15/86; C07; C07K; C07K14/00; C07K14/435; C07K14/575; C07K14/61; C07K14/795; C12N2710/00; C12N2710/00011; C12N2710/22011; C12N2710/22041; C12N2710/22043
Claims Analysis
Total Claims: 3
Claim 1
Independent Claim
159 words
A process for producing foreign proteins in eukaryotic cells comprising (a) cleaving SV40 at 0.14 map unit with BAM HI and 0.00 map unit with EcoRI to produce a SV-40 DNA fragment containing the origin of replication; (b) inserting said fragment into bacterial plasmid pBR322 together with pBR322 linkers between a single BAM HI site and a single EcoR1 site to produce pBR322-SV40; (c) after cloning cleaving pBR322-SV40 at the single BAM HI site and inserting a DNA fragment from a foreign gene therein; (d) cloning the resulting recombinant plasmid in E. coli; (e) remiving the plasmid pBR322 sequences by digesting the product with Hha I to generate SV-40 - foreign gene recombinant molecule containing a pBR322 linker; (f) treating said recombinant molecule with appropriate DNA ligase to cyclize the recombinant molecule, then using said recombinant molecule to infect African green monkey kidney cells together with a temperature sensitive tsA 239 SV40 DNA as the temperature-sensitive conditional lethal helper.
Claim 2
Dependent Claim
159 words
A process for producing foreign proteins in eukaryotic cells comprising (a) cleaving SV40 at 0.14 map unit with BAM HI and 0.00 map unit with EcoRI to produce a SV-40 DNA fragment containing the origin of replication; (b) inserting said fragment into bacterial plasmid pBR322 together with pBR322 linkers between a single BAM HI site and a single EcoR1 site to produce pBR322-SV40; (c) after cloning cleaving pBR322-SV40 at the single BAM HI site and inserting a DNA fragment from a foreign gene therein; (d) cloning the resulting recombinant plasmid in E. coli; (e) remiving the plasmid pBR322 sequences by digesting the product with Hha I to generate SV-40 - foreign gene recombinant molecule containing a pBR322 linker; (f) treating said recombinant molecule with appropriate DNA ligase to cyclize the recombinant molecule, then using said recombinant molecule to infect African green monkey kidney cells together with a temperature sensitive tsA 239 SV40 DNA as the temperature-sensitive conditional lethal helper.
Claim 3
Dependent Claim
26 words
The process in claim 1 in which the foreign gene fragment is one from the group consisting of: Mouse B MAJ globing and human growth hormone.
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) | 1989-07-07 | 1990-01-06 | 1990-07-08 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-07-07 | 1994-01-06 | 1994-07-08 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-07-07 | 1998-01-06 | 1998-07-08 | $7700.00 | Window Closed / Lapsed |