title: Isolation and characterization of replication-competent molecular DNA clones of HIV type 1 CRF01_AE with different coreceptor usages.
abstract: We have isolated replication-competent molecular clones of HIV-1 circulating recombinant form CRF01_AE with different coreceptor usages. After lambda phage cloning of unintegrated circular proviral DNAs derived from a CRF01_AE strain (HIV-1NH1), isolated in Japan, the infectious molecular clone, designated p93JP-NH1, was reconstituted. 93JP-NH1 showed an X4 and R5 phenotype in NP2 cell-based coreceptor utilization assays and exerted robust replication in human T cell lines, including MT2, M8166, and PM1 cells, whereas it propagated modestly in peripheral blood mononuclear cells. The CRF01_AE molecular clone with R5 phenotype (p93JP-NH2env) was then constructed by replacing the env gene of p93JP-NH1 with that of a nearly isogenic CRF01_AE R5 strain isolated from an epidemiologically linked case. The phylogeny and recombination break-point analysis confirmed that these clones shared an A/E recombinant structure similar to that of the prototype CRF01_AE strain, CM240. These replication-competent CRF01_AE molecular clones with different coreceptor usages would be useful tools for the study of CRF01_AE, one of the most prevalent strains in Asia.
authors.affiliation: Laboratory of Molecular Virology and Epidemiology, AIDS Research Center, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kusagawa
authors.forename: Hironori
authors.initials: H
authors.lastname: Sato
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Tomita
authors.forename: Masashi
authors.initials: M
authors.lastname: Tatsumi
authors.forename: Kayoko
authors.initials: K
authors.lastname: Kato
authors.forename: Kazushi
authors.initials: K
authors.lastname: Motomura
authors.forename: Rongge
authors.initials: R
authors.lastname: Yang
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Takebe
mesh_terms: D001483:Base Sequence; D015704:CD4 Antigens / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D004279:DNA, Viral; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D019713:Receptors, CCR5 / Q000235:genetics / Q000378:metabolism*; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid; D014779:Virus Replication
chemical_list: D015704:CD4 Antigens; D004279:DNA, Viral; D019713:Receptors, CCR5; D019718:Receptors, CXCR4