An infectious DNA clone of HIV type 1 subtype C

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 th e near future; however, there has been no report on the infectious DNA clon e 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 cell s, which are derived from HeLa-CD4-LTR-beta-gal (MAGI) cells and express CC R5, 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 re sulting plasmid was designated as pIndie-C1, The insert of pIndie-C1 was 96 80 bp in length and had an intact genomic organization with open reading fr ames of all structural, regulatory, and accessory proteins. Phylogenetic an alysis confirmed that the nucleotide sequence of pIndie-C1 is closely relat ed 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 widel y 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 patient s of India, but not of Japan. Expression of Nef and Vpr was also confirmed by immunoblotting.