Document 0736
 DOCN  M94A0736
 TI    Detection of HIV-1 tat, rev and nef mRNA at single cell level.
 DT    9412
 AU    Peng H; Haase AT; National Centre for HIV Virology Research, Institute
       of Medical; and Veterinary Science, Adelaide, South Australia.
 SO    Annu Conf Australas Soc HIV Med. 1993 Oct 28-30;5:104 (poster no. 59).
       Unique Identifier : AIDSLINE ASHM5/94348919
 AB    We have developed a strategy of in situ hybridisation using
       splice-junction probes which can specifically detect HIV-1 regulatory
       and structural gene mRNAs. This approach takes advantage of the fact
       that although tat, rev and nef share the same 5' splice donor, they
       differ in their use of the first splice acceptors. Eukaryotic expression
       vectors containing HIV-1 tat, rev, nef, env and gag genes were
       constructed and used to transfect Cos, Raji and H9 cells. The expression
       of HIV-1 genes on each transfectant was verified by indirect
       immunofluorescence. The specificity of the 35S-labelled splice junction
       probes was tested against all the transfectants under different
       stringency conditions. The conditions of in situ hybridisation were
       optimized, achieving the best specificity and sensitivity. This
       technique was successfully applied to detect HIV-1 tat, rev, nef, env
       and gag/genomic RNA in HIV-1 infected cells.
 DE    Cell Line  Genes, gag/GENETICS  Genes, nef/*GENETICS  Genes,
       rev/*GENETICS  Genes, tat/*GENETICS  Human  HIV-1/*GENETICS  In Situ
       Hybridization  RNA Splicing  RNA, Messenger/*GENETICS
       Transfection/GENETICS  MEETING ABSTRACT

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       protected by Copyright Law (Title 17, U.S.Code).