US4357421A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Synthetic gene coding for influenza hemagglutinin
Assignee: GD Searle LLC
•
Filing: 1980-03-24
•
Granted: 1982-11-02
•
Expires: 2000-03-24
Abstract
Abstract A process for the production of a synthetic gene for an influenza haemagglutinin which comprises: (a) subjecting isolated vRNA to the action of reverse transcriptase to produce a double-stranded RNA/DNA hybrid; (b) digesting the RNA component of the hybrid; (c) treating the residual DNA component of the hybrid to produce a double-stranded DNA molecule having one "hairpin" end; and (d) treating the "hairpin" structure with a single-strand specific nuclease to produce a bimolecular double-stranded copy of the vRNA.
Inventors
- 👤 John S. Emtage
- 👤 Norman H. Carey
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 20:41
CPC Classifications
C; C07; C07H; C07H21/00; A; A61; A61P; A61P31/00; A61P31/12; C07K; C07K14/00; C07K14/005; C12; C12P; C12P21/00; C12P21/02; A61K; A61K39/00; C12N; C12N2760/00; C12N2760/00011; C12N2760/16011; C12N2760/16111; C12N2760/16122
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
79 words
A process for the production of a synthetic gene for an influenza haemagglutinin comprising: (a) subjecting isolated influenza vRNA to the action of reverse transcriptase to produce a double-stranded RNA/DNA hybrid; (b) digesting the RNA component of the hybrid; (c) treating the residual DNA component of the hybrid to produce a double-stranded DNA molecule having one "hairpin" end; and (d) treating the "hairpin" structure with a single strand specific nuclease to produce a bimolecular double-stranded copy of the vRNA.
Claim 2
Dependent Claim
79 words
A process for the production of a synthetic gene for an influenza haemagglutinin comprising: (a) subjecting isolated influenza vRNA to the action of reverse transcriptase to produce a double-stranded RNA/DNA hybrid; (b) digesting the RNA component of the hybrid; (c) treating the residual DNA component of the hybrid to produce a double-stranded DNA molecule having one "hairpin" end; and (d) treating the "hairpin" structure with a single strand specific nuclease to produce a bimolecular double-stranded copy of the vRNA.
Claim 3
Dependent Claim
31 words
A process according to claim 1 wherein, in (a), a primer is used which is a deoxynucleotide sequence complementary to a known sequence in the 3' terminal sequence of the vRNA.
Claim 4
Dependent Claim
56 words
A process according to claim 1 wherein, in (a), the vRNA is treated with ATP:RNA adenyltransferase in the presence of Mg 2+ and/or Mn 2+ to produce a 3' polyriboadenylic acid extension of the vRNA and reverse transcription is carried out in the presence of a primer which is a homo-oligodeoxynucleotide complementary to the 3' extension.
Claim 5
Dependent Claim
32 words
A process according to claim 1 wherein, in (a), a pre-existing 3' terminal nucleotide sequence present in the vRNA is used as a primer resulting in self-priming in the reverse transcription reaction.
Claim 6
Dependent Claim
23 words
A process according to claim 1 wherein, in (b), the RNA component of the RNA/DNA hybrid is digested with an alkali or ribonuclease.
Claim 7
Dependent Claim
27 words
A process according to claim 1 wherein, in (c), the single-stranded DNA is converted into a double-stranded DNA "hairpin" structure with reverse transcriptase or a DNA polymerase.
Claim 8
Dependent Claim
16 words
A process according to claim 1 wherein, in (d), the single-strand specific nuclease is S1 nuclease.
Claim 9
Dependent Claim
15 words
A synthetic gene for an influenza haemagglutinin produced by the process according to claim 1.
Claim 10
Dependent Claim
33 words
A plasmid vector comprising, inserted therein, a synthetic gene according to claim 8, the plasmid reading in the correct phase for the expression of the gene and having a bacterial promoter upstream thereof.
Claim 11
Dependent Claim
19 words
A process for the transformation of a transformable cell comprising inserting therein a plasmid vector according to claim 9.
Claim 12
Dependent Claim
16 words
A process according to claim 10 wherein clones containing the haemagglutinin gene are identified and separated.
Claim 13
Dependent Claim
14 words
A cell comprising a plasmid vector produced by a process according to claim 9.
Claim 14
Dependent Claim
12 words
A cell according to claim 12 which is an E. Coli cell.
Claim 15
Dependent Claim
27 words
A process for the production of a protein by the expression of a synthetic gene for an influenza haemagglutinin comprising culturing a cell according to claim 12.
Claim 16
Dependent Claim
15 words
A plasmid vector according to claim 9 wherein the plasmid is of the pWT series.
Claim 17
Dependent Claim
19 words
A process for the transformation of a transformable cell comprising inserting therein a plasmid vector according to claim 15.
Claim 18
Dependent Claim
16 words
A process according to claim 16 wherein clones containing the haemagglutinin gene are identified and separated.
Claim 19
Dependent Claim
14 words
A cell comprising a plasmid vector produced by a process according to claim 15.
Claim 20
Dependent Claim
12 words
A cell according to claim 18 which is an E. Coli cell.
Claim 21
Dependent Claim
27 words
A process for the production of a protein by the expression of a synthetic gene for an influenza haemagglutinin comprising culturing a cell according to claim 16.
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) | 1985-11-01 | 1986-05-03 | 1986-11-02 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1989-11-01 | 1990-05-03 | 1990-11-02 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1993-11-01 | 1994-05-03 | 1994-11-02 | $7700.00 | Window Closed / Lapsed |