Purified pol DNA, a recombinant DNA construct for expression of HIV pol polypeptide sequences and a cell containing such construct
View Patent ↗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.
1. A recombinant DNA construct useful for the expression of a recombinant polypeptide in a cell containing the construct, the construct comprising control sequences which regulate transcription and translation of the recombinant polypeptide in the cell and a coding sequence regulated by the control sequences, wherein the coding sequence comprises a DNA sequence of at least about 27 bp of the pol open reading frame shown in FIG. 4 , characterized in that the DNA sequence encodes an antigenic pol polypeptide having a sequence within BstXI (3006) to NdeI (5131) which sites are set forth in FIG. 4 , which polypeptide sequence is immunologically non-cross-reactive with HTLV-I and HTLV-II and is reactive with HIV.
2. A recombinant DNA construct according to claim 1 which is useful for expression in a eukaryotic cell.
3. A recombinant DNA construct according to claim 1 which is useful for expression in a yeast cell.
4. A recombinant DNA construct according to claim 1 which is useful for expression in a bacterial cell.
5. A recombinant DNA construct according to claim 1 wherein the DNA encodes a complete pol polypeptide.
6. A cell comprising a recombinant DNA construct according to any one of claims 1 to 5 , wherein the cell expresses the antigenic HIV amino acid sequence and is free from other cells which do not express the antigenic HIV amino acid sequence.
7. A cell according to claim 6 wherein the recombinant DNA construct comprises a replication system recognized by the cell.
8. A cell according to claim 7 wherein the cell is eukaryotic.
9. A cell according to claim 8 which is a yeast cell.
10. A cell according to claim 9 which is a bacteria cell.
11. A recombinant antigenic pol HIV polypeptide produced by the construct of claim 2 expressed in an eukaryotic cell.
12. A recombinant antigenic pol HIV polypeptide produced by the construct of claim 3 expressed in a yeast cell.
13. A recombinant antigenic pol HIV polypeptide produced by the construct of claim 4 expressed in a bacterial cell.
14. A recombinant antigenic pol HIV polypeptide produced by the DNA construct of claim 1 .
15. A recombinant antigenic pol HIV polypeptide produced by the DNA construct of claim 5 .
16. A purified polynucleotide of at least about 20 bp of the pol open reading frame shown in FIG. 4 which encodes an antigenic pol polypeptide having a sequence within BstXI (3006) to NdeI (5131) which sites are set forth in FIG. 4 , which polypeptide sequence is immunologically non-cross-reactive with HTLV-I and HTLV-II and is reactive with HIV.