       Document 0452
 DOCN  M9460452
 TI    Molecular and biological characterization of simian immunodeficiency
       virus macaque strain 32H proviral clones containing nef size variants.
 DT    9404
 AU    Rud EW; Cranage M; Yon J; Quirk J; Ogilvie L; Cook N; Webster S; Dennis
       M; Clarke BE; Wellcome Research Laboratories, Department of Molecular
       Sciences,; Beckenham, Kent, U.K.
 SO    J Gen Virol. 1994 Mar;75 ( Pt 3):529-43. Unique Identifier : AIDSLINE
       GENBANK/D01065
 AB    The proviral genome of the 32H reisolate of simian immunodeficiency of
       macaques (SIVmac32H) has been cloned and sequenced. Including both long
       terminal repeats, it is 10277 base pairs in length and contains open
       reading frames for all known SIV genes (gag, pol, vif, vpx, vpr, tat,
       rev, env and nef). This is the first report of an infectious SIVmac
       molecular clone which contains no premature termination codons. Three
       molecular clones of SIVmac32H have been constructed differing in
       sequence only within their last 1.2 kb. Two of the molecular clones,
       SIVmac32H(pJ5) and SIVmac32H (pC8), differ in the nef coding region by
       an in-frame deletion of four amino acids in pC8 and two conservative
       amino acid changes; other nucleotide changes in the 3' LTR were not
       associated with known functionally critical motifs. The third clone,
       SIVmac32H(pB1), contains the last 1.2 kb of the SIVmac251 clone pBK28.
       The biological properties of virus produced after electroporation of
       these clones into C8166 cells has been assessed by infection of rhesus
       and cynomolgus macaques, time to seroconversion and by induction of
       cytopathic effects upon co-cultivation of infected rhesus peripheral
       blood lymphocytes with C8166 cells. The viruses obtained from these
       clones have identical growth kinetics in vitro but differ in their
       ability to persist in macaques. Macaques infected with pJ5 derived virus
       remain viraemic longer than macaques infected with pC8-derived virus.
       PCR analysis of circulating provirus indicates that the nef gene evolved
       over time in pJ5 virus-infected macaques, whereas late in infection in
       pC8 virus-infected macaques the nef gene remained invariant in sequence.
       These results support the observation that a nef deletion mutant of
       SIVmac239 lost its pathogenic potential and resulted in low-level
       viraemia when rhesus macaques were infected. Virus challenge pools for
       vaccine studies have been prepared for pJ5 using both human and monkey
       cell substrates and these stocks have been titrated both in vitro and in
       vivo. Virus has also been prepared from pC8 and titrated in vitro. This
       virus pool is being assessed as an attenuated live-virus vaccine in
       macaques. Since only virus originating from the SIVmac239 molecular
       clone is known to cause AIDS-like symptoms in rhesus macaques
       consistently, the SIVmac32H molecular clones should tell us more about
       which viral sequence features are important for the pathogenesis of
       AIDS.
 DE    Amino Acid Sequence  Animal  Base Sequence  Cell Line  Cloning,
       Molecular  Genes, nef/*GENETICS/IMMUNOLOGY  Macaca mulatta/*MICROBIOLOGY
       Molecular Sequence Data  Polymerase Chain Reaction
       Proviruses/*GENETICS/*IMMUNOLOGY  Simian Acquired Immunodeficiency
       Syndrome/IMMUNOLOGY  Support, Non-U.S. Gov't  SIV/GROWTH &
       DEVELOPMENT/*GENETICS/*IMMUNOLOGY  Tissue Culture  Viral Vaccines
       Virion  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

