Vector for expression of a polypeptide in a mammalian cell
AssigneeChiron Corporation
InventorsPaul A. Luciw, Dino Dina, Steven Rosenberg, Barbara S. Chapman, Richard M. Thayer, Nancy L. Haigwood
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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.
Claims24
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.
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.
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.
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.
Dependencies are inferred from the claim text.