US5688688A
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Vector for expression of a polypeptide in a mammalian cell
Assignee: Grifols Worldwide Operations Ltd
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Filing: 1994-08-10
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Granted: 1997-11-18
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Expires: 2014-08-10
Abstract
Abstract Polynucleotide sequences are provided for the diagnosis of the presence of retroviral infection in a human host associated with lymphadenopathy syndrome and/or acquired immune deficiency syndrome, for expression of polypeptides and use of the polypeptides to prepare antibodies, where both the polypeptides and antibodies may be employed as diagnostic reagents or in therapy, e.g., vaccines and passive immunization. The sequences provide detection of the viral infectious agents associated with the indicated syndromes and can be used for expression of antigenic polypeptides.
Inventors
- 👤 Paul A. Luciw
- 👤 Dino Dina
- 👤 Steven Rosenberg
- 👤 Barbara S. Chapman
- 👤 Richard M. Thayer
- 👤 Nancy L. Haigwood
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 07:43
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/005; A; A61; A61P; A61P31/00; A61P31/12; A61P31/14; A61P31/18; A61P37/00; A61P37/02; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/80; C12N15/81; C12N15/85; C12N9/00; C12N9/0004; C12N9/0089; C12Q; C12Q1/00; C12Q1/70; C12Q1/701; C12Q1/702; C12Q1/703; A61K; A61K38/00; C07K2319/00; C07K2319/40; C07K2319/70; C07K2319/735; C07K2319/74; C07K2319/75; C12N2740/00; C12N2740/00011; C12N2740/10011; C12N2740/16011; C12N2740/16111; C12N2740/16122; C12N2740/16211; C12N2740/16222; C12N2800/00; C12N2800/10; C12N2800/108; C12N2830/00; C12N2830/001; C12N2830/002; C12N2830/005; C12N2830/15; C12N2830/36; C12N2830/55; C12N2830/70; C12N2830/702; C12N2830/704; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/82; Y10S435/8215; Y10S435/948; Y10S435/974; Y10S435/975; Y10S530/00; Y10S530/82; Y10S530/826
Claims Analysis
Total Claims: 28
Claim 1
Independent Claim
99 words
A vector for expression of a polypeptide in a mammalian cell comprising a first polynucleotide sequence that comprises: a) an upstream SV40 origin of replication; b) a downstream SV40 polyadenylation region; and c) a transcription regulatory region from human cytomegalovirus immediate early region HCMV IE1, wherein the transcription regulatory region includes the first HCMV IE1 intron proximal to the 3' end of the HCMV IE1 promoter, is interposed between the SV40 origin of replication and the SV40 polyadenylation region, and is capable of directing the transcription of a polypeptide coding sequence operably linked downstream from the transcription regulatory region.
Claim 2
Dependent Claim
99 words
A vector for expression of a polypeptide in a mammalian cell comprising a first polynucleotide sequence that comprises: a) an upstream SV40 origin of replication; b) a downstream SV40 polyadenylation region; and c) a transcription regulatory region from human cytomegalovirus immediate early region HCMV IE1, wherein the transcription regulatory region includes the first HCMV IE1 intron proximal to the 3' end of the HCMV IE1 promoter, is interposed between the SV40 origin of replication and the SV40 polyadenylation region, and is capable of directing the transcription of a polypeptide coding sequence operably linked downstream from the transcription regulatory region.
Claim 3
Dependent Claim
27 words
The vector of claim 1, wherein the polynucleotide sequence further comprises a linker that comprises a restriction site for insertion of the coding region of a polypeptide.
Claim 4
Dependent Claim
13 words
The vector of claim 2, wherein the restriction site is a SalI site.
Claim 5
Dependent Claim
19 words
The vector of claim 1, wherein the SV40 polyadenylation region comprises the SV40 polyadenylation sequence present in plasmid pSV7d.
Claim 6
Dependent Claim
22 words
The vector of claim 1, wherein the SV40 origin of replication comprises the SV40 origin of replication sequence present in plasmid pSVT2.
Claim 7
Dependent Claim
10 words
The vector of claim 1, further comprising a selectable marker.
Claim 8
Dependent Claim
17 words
The vector of claim 5, wherein the selectable marker is a polynucleotide sequence that encodes ampicillin resistance.
Claim 9
Dependent Claim
12 words
The vector of claim 1, further comprising a bacterial origin of replication.
Claim 10
Dependent Claim
21 words
The vector of claim 1, wherein the polynucleotide sequence comprises the HCMV sequences present in plasmid pCMV6ARV120tpa, ATCC Accession No. 68249.
Claim 11
Dependent Claim
19 words
The vector of claim 2, further comprising a coding region that encodes a polypeptide, inserted at the restriction site.
Claim 12
Dependent Claim
32 words
The vector of claim 10, further comprising a region encoding a signal sequence effective in directing the secretion of the polypeptide encoded by the coding region, positioned upstream from the coding region.
Claim 13
Dependent Claim
19 words
The vector of claim 11, wherein the signal sequence is derived from the human tissue plasminogen activator leader sequence.
Claim 14
Dependent Claim
74 words
A vector produced by the process comprising linking together in an operative manner: a) a SV40 origin of replication; b) a SV40 polyadenylation region; and c) a transcription regulatory region from human cytomegalovirus immediate early region HCMV IE1, wherein said regulatory region includes the first HCMV IE1 intron proximal to the 3' end of the HCMV IE1 promoter and is capable of directing the transcription of a polypeptide coding sequence operably linked downstream therefrom.
Claim 15
Dependent Claim
74 words
A vector produced by the process comprising linking together in an operative manner: a) a SV40 origin of replication; b) a SV40 polyadenylation region; and c) a transcription regulatory region from human cytomegalovirus immediate early region HCMV IE1, wherein said regulatory region includes the first HCMV IE1 intron proximal to the 3' end of the HCMV IE1 promoter and is capable of directing the transcription of a polypeptide coding sequence operably linked downstream therefrom.
Claim 16
Dependent Claim
17 words
The vector of claim 13, wherein the vector is arranged in the same manner as plasmid pCMV6a.
Claim 17
Independent Claim
124 words
A method for producing a vector for expression of a polypeptide in a mammalian cell comprising: a) providing a first polynucleotide molecule that comprises a SV40 origin of replication; b) providing a second polynucleotide molecule that comprises a SV40 polyadenylation region; c) providing a third polynucleotide molecule that comprises a transcription regulatory region from human cytomegalovirus immediate early region HCMV IE1, wherein said regulatory region includes the first HCMV IE1 intron proximal to the 3' end of the HCMV IE1 promoter; and d) linking the SV40 origin of replication, the SV40 polyadenylation region and the regulatory region from HCMV IE1 together to form a vector that is capable of effecting the transcription of a polypeptide coding sequence operatively linked downstream from the regulatory region.
Claim 18
Independent Claim
124 words
A method for producing a vector for expression of a polypeptide in a mammalian cell comprising: a) providing a first polynucleotide molecule that comprises a SV40 origin of replication; b) providing a second polynucleotide molecule that comprises a SV40 polyadenylation region; c) providing a third polynucleotide molecule that comprises a transcription regulatory region from human cytomegalovirus immediate early region HCMV IE1, wherein said regulatory region includes the first HCMV IE1 intron proximal to the 3' end of the HCMV IE1 promoter; and d) linking the SV40 origin of replication, the SV40 polyadenylation region and the regulatory region from HCMV IE1 together to form a vector that is capable of effecting the transcription of a polypeptide coding sequence operatively linked downstream from the regulatory region.
Claim 19
Dependent Claim
29 words
A method for producing the vector of claim 1, comprising introducing the vector into a host cell and allowing the host cell to generate a plurality of said vectors.
Claim 20
Dependent Claim
37 words
An isolated nucleic acid molecule comprising an enhanced promoter, wherein the enhanced promoter comprises the human cytomegalovirus immediate early region HCMV IE1 promoter and the first intron proximate to the 3' end of the HCMV IE1 promoter.
Claim 21
Dependent Claim
37 words
An isolated nucleic acid molecule comprising an enhanced promoter, wherein the enhanced promoter comprises the human cytomegalovirus immediate early region HCMV IE1 promoter and the first intron proximate to the 3' end of the HCMV IE1 promoter.
Claim 22
Dependent Claim
21 words
The nucleic acid molecule of claim 17, wherein the promoter region is derived from a subclone of human cytomegalovirus (Towne strain).
Claim 23
Dependent Claim
43 words
A vector for expression of a polypeptide in a mammalian cell, comprising the nucleic acid molecule of claims 17, wherein the nucleic acid molecule is capable of directing the transcription of a polypeptide coding sequence operably linked downstream of the nucleic acid molecule.
Claim 24
Dependent Claim
19 words
The vector of claim 19, further comprising an origin of replication operably linked upstream of the nucleic acid molecule.
Claim 25
Dependent Claim
18 words
The vector of claim 19, further comprising a polyadenylation region operably linked downstream of the nucleic acid molecule.
Claim 26
Dependent Claim
65 words
A vector for expression of a polypeptide in a mammalian cell, comprising: a) an upstream origin of replication; b) a downstream polyadenylation region; and c) the nucleic acid molecule of claim 17 interposed between the origin of replication and the polyadenylation region, wherein the enhanced promoter region is capable of directing the transcription of a polypeptide coding sequence operably linked downstream from the promoter region.
Claim 27
Dependent Claim
22 words
A method for constructing the vector of claim 19, comprising operatively linking together the nucleic acid molecule and the polypeptide coding sequence.
Claim 28
Dependent Claim
34 words
A method for producing the vector constructed in claim 23, comprising introducing the vector into a host cell that is capable of replicating the vector and allowing the host cell to replicate the vector.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2000-11-17 | 2001-05-19 | 2001-11-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2004-11-17 | 2005-05-19 | 2005-11-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2008-11-17 | 2009-05-19 | 2009-11-18 | $7700.00 | Window Closed / Lapsed |