Molecular cloning of HIV-1 from immortalized cell lines

摘要

Disclosed is the molecular cloning of HTLV-III, the adult leukemia and acquired immune deficiency syndrome (AIDS) virus. Clone BH10 contains a 9.0 Kb viral insert constituting the entire HTLV-III genome. Clones BH8 and BH5 contain viral inserts of 5.5 Kb and 3.5 Kb, respectively. These clones are suitable for the development of diagnostic and therapeutic measures for AIDS, as well as use as probes for the detection of AIDS. By scientific convention, HTLV-III, referred to herein also as HIV, has been renamed as HIV-1.

主权项

1. A composition comprising a duplex formed between a Human Immunodeficiency Virus Type-1 (HIV-1) nucleic acid and a single-stranded nucleic acid of at least 18 contiguous nucleotides comprising a nucleotide sequence selected from the group consisting of:
(i) a nucleotide sequence of either strand of viral DNA from lambda bacteriophage λ-HXB2 having ATCC Accession No. 40231; (ii) a nucleotide sequence of either strand of viral DNA from lambda bacteriophage λ-HXB3 having ATCC Accession No. 40232; (iii) a nucleotide sequence of either strand of viral DNA from clone BH10 having ATCC Accession No. 40125; (iv) a nucleotide sequence of either strand of viral DNA from clone BH5 having ATCC Accession No. 40126; (v) a nucleotide sequence of either strand of viral DNA from clone BH8 having ATCC Accession No. 40127; (vi) a nucleotide sequence of either strand of viral DNA from clone pHXB3 having ATCC Accession No. 67081; (vii) a nucleotide sequence of either strand of viral DNA from clone pHXB-2D having ATCC Accession No. 67082; (viii) a nucleotide sequence of either strand of viral DNA from E. coli clone X10-1 having ATCC Accession No. 67083; and (ix) an HIV-1 nucleotide sequence from the H9/HTLV-III cell line having ATCC Accession No. CRL 8543;
wherein the single-stranded nucleic acid does not form a duplex with HTLV-I and HTLV-II nucleic acids under conditions of stringency for hybridization under which the single-stranded nucleic acid forms a duplex with the HIV-1 nucleic acid;wherein the duplex is outside of a mammalian cell and outside of an HIV-1 particle;wherein the duplex comprises a double-stranded region of at least 18 contiguous nucleotides and a single-stranded region on either side of the double-stranded region that is longer than the double-stranded region; andwherein the single-stranded nucleic acid comprises a label.