title: An infectious DNA clone of HIV type 1 subtype C.
abstract: Among the 10 subtypes of the M group of human immunodeficiency virus type 1, subtype C is the most prevalent in India and may dominate worldwide in the near future; however, there has been no report on the infectious DNA clone of this subtype. We have isolated an infectious DNA clone of the 93IN101 strain of HIV-1 subtype C, which was isolated in India in 1993. MAGIC5 cells, which are derived from HeLa-CD4-LTR-beta-gal (MAGI) cells and express CCR5, were inoculated with the 93IN101 strain of HIV-1 subtype C. The genomic DNA of the infected cells was used as a template for amplification of the HIV-1 genome. The genome DNA obtained was subcloned into pBR322, and the resulting plasmid was designated as pIndie-C1. The insert of pIndie-C1 was 9680 bp in length and had an intact genomic organization with open reading frames of all structural, regulatory, and accessory proteins. Phylogenetic analysis confirmed that the nucleotide sequence of pIndie-C1 is closely related to those of HIV-1 subtype C isolated in India. Transfection of pIndie-C1 into 293T cells yielded as much virus as did pNL432, one of the most widely used HIV DNA clones. The recovered Indie-C1 virus infected MAGIC5 but not the parent MAGI cells, indicating that Indie-C1 is CCR5 tropic. Expressed Env protein was reacted efficiently with the sera of HIV-1-infected patients of India, but not of Japan. Expression of Nef and Vpr was also confirmed by immunoblotting.
authors.affiliation: Department of Pathology, Research Institute, International Medical Center of Japan, Toyama, Tokyo.
authors.forename: N
authors.initials: N
authors.lastname: Mochizuki
authors.forename: N
authors.initials: N
authors.lastname: Otsuka
authors.forename: K
authors.initials: K
authors.lastname: Matsuo
authors.forename: T
authors.initials: T
authors.lastname: Shiino
authors.forename: A
authors.initials: A
authors.lastname: Kojima
authors.forename: T
authors.initials: T
authors.lastname: Kurata
authors.forename: K
authors.initials: K
authors.lastname: Sakai
authors.forename: N
authors.initials: N
authors.lastname: Yamamoto
authors.forename: S
authors.initials: S
authors.lastname: Isomura
authors.forename: T N
authors.initials: TN
authors.lastname: Dhole
authors.forename: Y
authors.initials: Y
authors.lastname: Takebe
authors.forename: M
authors.initials: M
authors.lastname: Matsuda
authors.forename: M
authors.initials: M
authors.lastname: Tatsumi
mesh_terms: D015686:Gene Products, env / Q000378:metabolism; D015693:Gene Products, nef / Q000378:metabolism; D016336:Gene Products, vpr / Q000378:metabolism; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D007194:India; D008969:Molecular Sequence Data; D010641:Phenotype; D010802:Phylogeny; D019713:Receptors, CCR5 / Q000378:metabolism; D017422:Sequence Analysis, DNA; D054311:nef Gene Products, Human Immunodeficiency Virus; D054325:vpr Gene Products, Human Immunodeficiency Virus
chemical_list: D015686:Gene Products, env; D015693:Gene Products, nef; D016336:Gene Products, vpr; D019713:Receptors, CCR5; D054311:nef Gene Products, Human Immunodeficiency Virus; D054325:vpr Gene Products, Human Immunodeficiency Virus