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Titolo:
A NONRADIOACTIVE MICROTITRE PLATE REVERSE-TRANSCRIPTASE (RT) ASSAY, BASED ON IMMOBILIZED TEMPLATE, FOR SCREENING OF RT ACTIVITY INHIBITORS AND EVALUATION OF THEIR MODE OF ACTION
Autore:
SHAO X; EKSTRAND DHL; BHIKHABHAI R; KALLANDER CFR; GRONOWITZ JS;
Indirizzi:
UNIV UPPSALA,DEPT MED GENET,RES UNIT REPLICAT ENZYMOL,BMC,BOX 584 S-75123 UPPSALA SWEDEN UNIV UPPSALA,DEPT MED GENET,RES UNIT REPLICAT ENZYMOL,BMC S-75123 UPPSALA SWEDEN PHARMACIA BIOTECH S-75182 UPPSALA SWEDEN CAVIDI TECH S-75183 UPPSALA SWEDEN CHINESE ACAD MED SCI,INST MED BIOTECHNOL,DEPT VIROL BEIJING 100050 PEOPLES R CHINA
Titolo Testata:
Antiviral chemistry & chemotherapy
fascicolo: 2, volume: 8, anno: 1997,
pagine: 149 - 159
SICI:
0956-3202(1997)8:2<149:ANMPR(>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
IMMUNODEFICIENCY-VIRUS INFECTIONS; ANTIVIRAL THERAPY; AZIDOTHYMIDINE TRIPHOSPHATE; RESISTANCE; TYPE-1; 3'-AZIDO-3'-DEOXYTHYMIDINE; PURIFICATION; SENSITIVITY; ZIDOVUDINE; MUTATIONS;
Keywords:
REVERSE TRANSCRIPTASE; RT ASSAYS; RT INHIBITORS; ANTIVIRALS; CARRIER BOUND TEMPLATE-PRIMERS; HIV/AIDS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
24
Recensione:
Indirizzi per estratti:
Citazione:
X. Shao et al., "A NONRADIOACTIVE MICROTITRE PLATE REVERSE-TRANSCRIPTASE (RT) ASSAY, BASED ON IMMOBILIZED TEMPLATE, FOR SCREENING OF RT ACTIVITY INHIBITORS AND EVALUATION OF THEIR MODE OF ACTION", Antiviral chemistry & chemotherapy, 8(2), 1997, pp. 149-159

Abstract

A new sensitive colorimetric reverse transcriptase (RT) activity assay utilizing a 96-well microtitre plate format, with solid phase-conjugated polyadenylic acid (prA), was investigated for simple analyses of the RT inhibiting capacity and mode of action of various substances. Three different technical procedures using the assay were evaluated: (i) direct IC50 determinations with various substances, using four different combinations of primer and dNTP amounts; (ii) analyses of the capacity of the substances to interfere with the binding of RT to template or template-primer (BIC50); (iii) analyses of the capacity of the substances to destroy the template-primer in presence or absence of RT (TDC50). The assay was found to be useful for all three purposes using small amounts of recombinant RT. In the IC50 analyses, the test substances gave values similar to those reported for soluble RT assays, and the values varied in accordance with their known mode of action in relation to the combination of primer and dNTP amount used. Only one of the substances, prG, in addition to DNA and RNA gave true RT binding inhibition. The template destruction assay showed that chain terminatingsubstances gave destruction at low inhibitor concentrations. Furthermore, this destruction was RT-dependent, in contrast to the destructionobtained with substances that can base-pair with the template or primer. For optimum information on mode of action of a given substance allthree assay procedures should be used. The use of the assay in relation to the screening and analyses of new RT inhibitory substances and characterization of RT in primary isolates or plasma is discussed.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 31/10/20 alle ore 00:03:16