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M94A0736.TXT
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1994-10-21
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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
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).