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Titolo:
CHARACTERIZATION OF FLUORESCEIN-OLIGONUCLEOTIDE CONJUGATES AND MEASUREMENT OF LOCAL ELECTROSTATIC POTENTIAL
Autore:
SJOBACK R; NYGREN J; KUBISTA M;
Indirizzi:
CHALMERS UNIV TECHNOL,DEPT BIOCHEM & BIOPHYS S-41390 GOTHENBURG SWEDEN CHALMERS UNIV TECHNOL,DEPT BIOCHEM & BIOPHYS S-41390 GOTHENBURG SWEDEN
Titolo Testata:
Biopolymers
fascicolo: 7, volume: 46, anno: 1998,
pagine: 445 - 453
SICI:
0006-3525(1998)46:7<445:COFCAM>2.0.ZU;2-0
Fonte:
ISI
Lingua:
ENG
Soggetto:
RESONANCE ENERGY-TRANSFER; DOUBLE-STRANDED DNA; SPECTRAL-ANALYSIS; HYBRIDIZATION; ABSORPTION;
Keywords:
FLUORESCEIN-OLIGONUCLEOTIDE CONJUGATES; LOCAL ELECTROSTATIC POTENTIAL; LOCAL PH; PROTOLYTIC EQUILIBRIUM; FITC FLUORESCENCE ENERGY TRANSFER; DYE LABELED OLIGONUCLEOTIDES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
33
Recensione:
Indirizzi per estratti:
Citazione:
R. Sjoback et al., "CHARACTERIZATION OF FLUORESCEIN-OLIGONUCLEOTIDE CONJUGATES AND MEASUREMENT OF LOCAL ELECTROSTATIC POTENTIAL", Biopolymers, 46(7), 1998, pp. 445-453

Abstract

The properties of fluorescein are substantially altered upon conjugation to nucleic acids, affecting not only the molar absorptivities and fluorescence quantum yields but also the protolytic equilibrium constant and fluorescence lifetimes. Around neutral pH, the fluorescein moiety is present as both mono- and dianion, and the pK(a) relating them is increased from 6.43 for free fluorescein to about 6.90 for fluorescein attached to both single- and double-stranded oligonucleotides of atleast 12 bases/base pairs. This difference reflects the local electrostatic potential around the nucleic acid, which is calculated to -28 mV. The molar absorptivities and spectral responses of the conjugated fluorescein protolytic species are also determined from which the concentrations of fluorescein-oligonucleotide conjugates can be calculated by assuming: 8(484) = 62000/[1 + 10(-(pH-6.90))] + 12000/[1 + 10((pH-690))] (M-1 cm(-1)). The fluorescence quantum yield of the conjugates depends, in a complex way, on temperature, environment and oligonucleotide length, sequence and conformation, and must be determined for eachexperimental situation. (C) 1998 John Wiley & Sons, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 19/09/20 alle ore 08:40:05