Detection of HIV-1 tat, rev and nef mRNA at single cell level.

Important note: Information in this article was accurate in 1994. The state of the art may have changed since the publication date.

Detection of HIV-1 tat, rev and nef mRNA at single cell level.

Annu Conf Australas Soc HIV Med. 1993 Oct 28-30;5:104 (poster no. 59). Unique Identifier : AIDSLINE ASHM5/94348919
Peng H; Haase AT; National Centre for HIV Virology Research, Institute of Medical; and Veterinary Science, Adelaide, South Australia.

Abstract: 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.

Keywords: 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 ABSTRACT
941230
M94C4308

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