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1994-08-27
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Document 0657
DOCN M9480657
TI Development of a human immunodeficiency virus-1 in vitro DNA synthesis
system to study reverse transcriptase inhibitors.
DT 9410
AU Borroto-Esoda K; Boone LR; Division of Virology, Wellcome Research
Laboratories, Burroughs; Wellcome Co., Research Triangle Park, North
Carolina 27709.
SO Antiviral Res. 1994 Apr;23(3-4):235-49. Unique Identifier : AIDSLINE
MED/94317913
AB A Human immunodeficiency virus type-1 endogenous reverse transcriptase
reaction was developed as an in vitro assay to study the inhibition of
reverse transcription by antiviral compounds. Conditions were
established for producing genomic length (-) strand DNA in high yields
and measuring the inhibition of this transcript as the assay endpoint.
In addition to genomic length (-) strand DNA, a novel segmented (-)
strand product composed of a 6.0 kb reverse transcript of the 5' 2/3 of
the viral RNA genome and a 3.5 kb reverse transcript of the 3' 1/3 was
observed. The most prominent (+) strand product was the size expected
for plus-strong stop DNA. Additional minor (+) strand species were also
observed. The triphosphate form of the nucleoside analog inhibitor
3'-azido-3'-deoxythymidine (RETROVIR, Zidovudine, AZT) and BI-RG-587
(nevirapine), a non nucleoside inhibitor, were used to demonstrate the
utility of the endogenous system for the analysis of reverse
transcriptase inhibitors. In a standard reaction, synthesis of genomic
length DNA was 50% inhibited by 0.1 microM AZTTP and 0.1 microM
nevirapine.
DE DNA, Viral/*BIOSYNTHESIS/CHEMISTRY Hydrogen-Ion Concentration
HIV-1/*ENZYMOLOGY/METABOLISM Pyridines/PHARMACOLOGY Reverse
Transcriptase/*ANTAGONISTS & INHIB/METABOLISM Thymine
Nucleotides/PHARMACOLOGY Time Factors Zidovudine/ANALOGS &
DERIVATIVES/PHARMACOLOGY JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).