Molecular cloning and biological characterization of full-length HIV-1 subtype C from Botswana.
Human immunodeficiency virus type 1 (HIV-1) subtype C is now responsible for more than half of all HIV-1 infections in the global epidemic and for the high levels of HIV-1 prevalence in southern Africa. To facilitate studies of the biological nature and the underlying molecular determinants of this virus, we constructed eight full-length proviral clones from two asymptomatic and three AIDS patients infected with HIV-1 subtype C from Botswana. Analysis of viral lysates showed that Gag, Pol, and Env structural proteins were present in the virions. In four clones, the analysis suggested inefficient envelope glycoprotein processing. Nucleotide sequence analysis of the eight clones did not reveal frameshifts, deletions, premature truncations, or translational stop codons in any structural, regulatory, or accessory genes. None of the subtype C clones were replication competent in donor peripheral blood mononuclear cells (PBMCs), macrophages, Jurkat(tat) cells, or U87. CD4.CCR5 cells. However, infection by two clones could be rescued by complementation with a functional subtype C envelope clone, resulting in a productive infection of PBMCs, macrophages, and U87. CD4.CCR5 cells.
Ndung'u
T
T
Harvard AIDS Institute, Harvard School of Public Health, Boston, Massachusetts 02115, USA.
Renjifo
B
B
Novitsky
V A
VA
McLane
M F
MF
Gaolekwe
S
S
Essex
M
M
D000163:Acquired Immunodeficiency Syndrome / Q000821:virology; D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D001902:Botswana; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D016384:Consensus Sequence; D005260:Female; D015686:Gene Products, env / Q000737:chemistry / Q000235:genetics*; D005910:Glioma; D015658:HIV Infections / Q000821:virology; D015497:HIV-1 / Q000145:classification* / Q000235:genetics* / Q000502:physiology; D006801:Humans; D019169:Jurkat Cells; D008214:Lymphocytes / Q000821:virology; D008264:Macrophages / Q000821:virology; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D014779:Virus Replication / Q000235:genetics
D015686:Gene Products, env