The Complete Nucleotide Sequence of a Pathogenic Molecular Clone of Simian Immunodeficiency Virus

Abstract

The complete nucleotide sequence of an infectious clone of simian immunodeficiency virus of macaques, SIVmac239, has been determined. Virus produced from this molecular clone causes AIDS in rhesus monkeys in a time frame suitable for laboratory investigation. The proviral genome including both long terminal repeats is 10,279 base pairs in length and contains open reading frames for gag, pol, vif, vpr, vpx, tat, rev, and env. The nef gene contains an in-frame premature stop after the 92nd codon. At the nucleotide level, SIVmac239 is closely related to SIVmac251 (98%) and SIVmac142 (96%). It will not be possible to test which features of the viral sequence are critical molecular determinants for the pathogenesis of AIDS.

No reactivation of attenuated immunodeficiency viruses in rhesus macaques after vaccinia virus-induced immune activation.

“…Similar effects on virus replication and pathogenicity have been found in monkeys infected with SHIV. In our experiment, Kuwata et al (1995) ; b, Kuwata et al (1996) ; c, Rud et al (1994) ; d, Cranage et al (1990) ; e, Regier & Desrosiers (1990). † Duration of immunodeficiency virus infection prior to VV super-infection.…”

Simian-Human Immunodeficiency Virus Escape from Cytotoxic T-Lymphocyte Recognition at a Structurally Constrained Epitope

“…gag mutants T47I, I71V, and T47I/I71V were created by PCR mutagenesis with a QuikChange kit (Stratagene) and p239SpSp5Ј as the template; p239SpSp5Ј was obtained through the AIDS Research and Reference Reagent Program, Division of AIDS, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), from Ronald Desrosiers (19,28). Oligonucleotide primers T47IF (5Ј-CAGGCACTGTCAGAAGGTTG CATCCCCTATGACATTAATCAGATGTTAAATTG-3Ј; nt 1831 to 1883) and T47IR (5Ј-CAATTTAACATCTGATTAATGTCATAGGGGATGCAACCTT CTGACAGTGCCTG-3Ј; nt 1831 to 1883) were used to change Thr (ACC) to Ile (ATC).…”

“…Plasmid pSHIV⌬envCAT and the envelope plasmids were kindly provided by J. Sodroski (Dana-Farber Cancer Institute, Boston, Mass.). Recombinant SIVmac239 viruses were produced as previously described (19,28). Briefly, 5 g of each proviral half was digested with SphI, phenol-chloroform extracted, ethanol precipitated, and ligated.…”

Effects of 6-Chloro-2',3'-Dideoxyguanosine (6-Cl-ddG) in Surface Lymph Nodes of Rhesus Monkeys (Macaca mulatta) Chronically Infected with Simian Immunodeficiency Virus (SIVmac239).

“…We used two juvenile rhesus monkeys (Mm9408 and Mm9421) that were intravenously inoculated with 100 TCID 50 of SIVmac239 [19] 12 months before being used in our experiments. Although these monkeys manifested lymphoid adenopathy, p27 gag antigen in their plasma was undetectable before the experiments.…”

“…Preparation of digoxigenin-labeled RNA probes: Portions of gag, pol, and nef genes of SIVmac239 [19] were inserted into pSPT19 and pSPT18 (Boehringer Mannheim), and each linearized product was used as a template for antisense and sense probes, respectively. All probes were generated from T7 promoter in the presence of digoxigenin-labeled UTP with a DIG RNA Labeling Kit (Boehringer Mannheim) according to the manufacturer's instruction.…”