Recombinant DNA constructs for GAG polypeptides and polypeptides produced thereby
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 gag open reading frame shown in FIG. 4 , characterized in that the DNA sequence encodes an antigenic gag HIV amino acid sequence which amino acid sequence is immunologically non-cross-reactive with HTLV-I and HTLV-II.
2. A recombinant DNA construct according to claim 1 which comprises a replication system functional in a eukaryotic cell.
3. A recombinant DNA construct according to claim 1 which comprises a replication system functional in a yeast cell.
4. A recombinant DNA construct according to claim 1 which comprises a replication system functional in a bacterial cell.
5. A recombinant DNA construct according to claim 1 wherein the antigenic amino acid sequence comprises p18 gag.
6. A recombinant DNA construct according to claim 1 wherein the antigenic amino acid sequence comprises p25 gag.
7. A recombinant DNA construct according to claim 1 wherein the antigenic amino acid sequence comprises a fusion protein of p18 gag and p25 gag.
8. A recombinant DNA construct according to any one of claims 1 - 4 wherein the antigenic amino acid sequence comprises a complete gag sequence.
9. A cell comprising a recombinant DNA construct according to any one of claims 1 to 4 , 5 , 6 , or 7 , 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.
10. A recombinant antigenic gag HIV polypeptide produced by the construct of claim 2 expressed in an eukaryotic cell.
11. A recombinant antigenic gag HIV polypeptide produced by the construct of claim 3 expressed in a yeast cell.
12. A recombinant antigenic gag HIV polypeptide produced by the construct of claim 4 expressed in a bacterial cell.
13. A recombinant antigenic gag HIV polypeptide produced by the DNA construct of claim 5 .
14. A recombinant antigenic gag HIV polypeptide produced by the DNA construct of claim 6 .
15. A recombinant antigenic gag HIV polypeptide produced by the DNA construct of claim 7 .
16. A recombinant antigenic gag HIV polypeptide produced by the DNA construct of claim 8 .
17. A purified polynucleotide of at least about 27 bp of the gag open reading frame shown in FIG. 4 which encodes an antigenic gag HIV amino acid sequence which amino acid sequence is immunologically non-cross-reactive with HTLV-1 and HTLV-II.
18. The polynucleotide of claim 17 wherein the antigenic amino acid sequence comprises p18 gag.
19. The polynucleotide of claim 17 wherein the antigenic amino acid sequence comprises p25 gag.
20. The polynucleotide of claim 17 wherein the antigenic amino acid sequence comprises a fusion protein of p18 gag and p25 gag.
21. The polynucleotide of claim 17 wherein the antigenic amino acid sequence comprises a complete gag sequence.