US4332892A
Expired / Lapsed
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Protein synthesis
Assignee: Harvard University
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Filing: 1980-01-10
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Granted: 1982-06-01
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Expires: 2000-01-10
Abstract
Abstract This invention is a process to produce specific proteins coded for by eukaryotic (or prokaryotic) DNA in bacteria. The invention, which uses recombinant DNA techniques, produces proteins in their natural, functional state unencumbered by extraneous peptides.
Inventors
- 👤 Mark Ptashne
- 👤 Gail D. Lauer
- 👤 Thomas M. Roberts
- 👤 Keith C. Backman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 02:09
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/70; C12N15/72
Claims Analysis
Total Claims: 7
Claim 1
Independent Claim
132 words
The method of producing native, unfused prokaryotic or eukaryotic protein in bacteria which comprises inserting into a bacterial plasmid a gene for a prokaryotic or eukaryotic protein and a portable promoter consisting of a DNA fragment containing a Shine-Dalgarno sequence and a transcription initiation site recognized by RNA polymerase and containing no protein translational start site, said promoter being inserted upstream from a protein ATG translational start site of said gene, at a position to obtain production of said protein, to form a fused gene having said Shine-Dalgarno sequence, said transcription initiation site, and the ATG signalling the start point of translation, inserting said plasmid into said bacteria to transform said bacteria with said plasmid containing said fused gene, and culturing the transformed bacteria to produce said unfused prokaryotic or eukaryotic protein.
Claim 2
Independent Claim
132 words
The method of producing native, unfused prokaryotic or eukaryotic protein in bacteria which comprises inserting into a bacterial plasmid a gene for a prokaryotic or eukaryotic protein and a portable promoter consisting of a DNA fragment containing a Shine-Dalgarno sequence and a transcription initiation site recognized by RNA polymerase and containing no protein translational start site, said promoter being inserted upstream from a protein ATG translational start site of said gene, at a position to obtain production of said protein, to form a fused gene having said Shine-Dalgarno sequence, said transcription initiation site, and the ATG signalling the start point of translation, inserting said plasmid into said bacteria to transform said bacteria with said plasmid containing said fused gene, and culturing the transformed bacteria to produce said unfused prokaryotic or eukaryotic protein.
Claim 3
Dependent Claim
14 words
The method as claimed in claim 1 in which said bacteria is E. coli.
Claim 4
Independent Claim
24 words
The method as claimed in claims 1 or 2 in which said portable promoter is the product of restriction endonuclease digestion of an operon.
Claim 5
Dependent Claim
25 words
The method claimed in claim 2 in which said portable promoter is the product of restriction endonuclease digestion of the lac operon of E. coli.
Claim 6
Independent Claim
48 words
The method as claimed in claims 1 or 2 in which said gene insertion comprises cloning said gene into said plasmid, adjusting the spacing between said inserted gene and a preceding unique restriction site by treating with a nuclease, and cloning said portable promoter into said restriction site.
Claim 7
Dependent Claim
57 words
The method as claimed in claim 2 in which said gene insertion comprises cloning said gene into said plasmid, adjusting the spacing between said gene and a preceding unique restriction site by treating with a nuclease, and cloning into said restriction site a portable promoter formed by the endonuclease digestion of the lac operon of E. coli.
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) | 1985-05-31 | 1985-11-30 | 1986-06-01 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1989-05-31 | 1989-11-30 | 1990-06-01 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1993-05-31 | 1993-11-30 | 1994-06-01 | $7700.00 | Window Closed / Lapsed |