A quantitative assay for HIV DNA integration in vivo.
Early steps of infection by HIV-1 involve entry of the viral core into cells, reverse transcription to form the linear viral DNA, and integration of that DNA into a chromosome of the host. The unintegrated DNA can also follow non-productive pathways, in which it is circularized by recombination between DNA long-terminal repeats (LTRs), circularized by ligation of the DNA ends or degraded. Here we report quantitative methods that monitor formation of reverse transcription products, two-LTR circles and integrated proviruses. The integration assay employs a novel quantitative form of Alu-PCR that should be generally applicable to studies of integrating viruses and gene transfer vectors.
Butler
S L
SL
Infectious Disease Laboratory, The Salk Institute for Biological Studies, La Jolla, California, USA.
Hansen
M S
MS
Bushman
F D
FD
D019380:Anti-HIV Agents / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D004279:DNA, Viral / Q000235:genetics*; D005453:Fluorescence; D015497:HIV-1 / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D006801:Humans; D016133:Polymerase Chain Reaction; D016662:Virus Integration / Q000187:drug effects; D014779:Virus Replication
D019380:Anti-HIV Agents; D017931:DNA Primers; D004279:DNA, Viral