Document 0927
 DOCN  M9540927
 TI    Quantitative and discriminative detection of individual HIV-1 mRNA
       subspecies by an RNAse mapping assay.
 DT    9504
 AU    Mary C; Telles JN; Cheynet V; Oriol G; Mallet F; Mandrand B; Verrier B;
       Unite mixte CNRS/Biomerieux, Lyon, France.
 SO    J Virol Methods. 1994 Aug;49(1):9-23. Unique Identifier : AIDSLINE
       MED/95130648
 AB    HIV-1 genes are expressed through the complex splicing of a single mRNA
       precursor leading to three mRNA classes: unspliced, singly-spliced and
       multiply-spliced. Each class may include several mRNA species
       specifically encoding one or two HIV-1 proteins. Northern blotting and
       RT-PCR are the techniques currently used to analyse HIV-1 mRNA
       expression. Northern blotting allows quantitative detection of these
       three classes of viral RNA but does not discriminate between individual
       RNA species. RT-PCR allows discrimination between different species but
       does not provide a quantitative analysis. Here, we describe an
       application of an RNAse mapping assay which gives both quantitative and
       discriminative HIV-1 RNA detection. A radiolabeled probe overlapping the
       major splicing sites of HIV-1 used for the generation of HIV-1 mRNA
       subspecies was synthesized. This probe protects differential sizes of
       these species, allowing discrimination between them. We investigated the
       RNA expression pattern in high titer HIV-1 producing cells. The
       HIV-1-specific probe allowed the detection of multiply-spliced vpr, rev
       and nef mRNAs, singly-spliced env mRNA and unspliced genomic RNA. With
       its discriminative and quantitative properties, this application is
       particularly convenient for the investigation of HIV-1 mRNA expression
       during the course of HIV-1 infections.
 DE    Base Sequence  Blotting, Northern  Cell Line  Cloning, Molecular
       Comparative Study  DNA Primers/GENETICS  DNA, Viral/GENETICS  Genes,
       Viral  Genetic Vectors  Human  HIV-1/*GENETICS  Molecular Sequence Data
       Nucleic Acid Hybridization  Polymerase Chain Reaction  *Ribonucleases
       RNA, Messenger/*ANALYSIS/*GENETICS  RNA, Viral/*ANALYSIS/*GENETICS
       Support, Non-U.S. Gov't  Virology/*METHODS  JOURNAL ARTICLE

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