A bstractThe pH-induced helix-coil transition of DNA and its com plexes w ith EtB r is carried out at acidic pH in a wide interval of change of concentration ratio of EtBr/DNA. The binding isotherm s of EtBr on double and single-stranded DNA at pH = 7.0 and pH = 3.0 (t = 25 o C) are obtained by absorption and fluorim etric m ethods. Binding constants (K) and num ber of bases (n), corresponding to one binding site were determ ined. Non fluorescent "strong" com plex w ith ds-DNA at pH = 7.0 and t = 25 o C as well as "strong" and "weak" complexes with ss-DNA at pH = 3.0 and t = 25 o C are revealed.
The binding of Hoechst 33258 with DNA at various ionic strengths of solution and different ligand concentrations has been investigated. Existence of more than one type of interactions of Hoechst 33258 with DNA has been revealed, which were very sensitive to the ionic strength. Hoechst 33258 doesn't show specificity to AT sequences of DNA at low ionic strength. High affinity binding mode becomes obvious at high ionic strength. The values of binding constants and binding site sizes for revealed strong and weak interactions have been determined.
Os parâmetros de ligação de brometo de etídio (EtBr) com DNA em diversas proporções GC/AT foram determinados usando espectroscopia de absorção e fluorescência. Nossos dados experimentais demonstram claramente a co-existência de tipos de ligação "forte" fluorescente e não-fluorescente em baixas concentrações de EtBr. O complexo fluorescente corresponde ao modelo normal intercalante. O complexo não-fluorescente refere-se à ligação semi-intercalante de EtBr. A constante de ligação (K) e o número de pares de bases correspondentes ao sítio de ligação (n) dos tipos fluorescente (K f e n f ) e não-fluorescente (K nf , n nf ) de interações foram determinados. O tamanho médio do sítio de ligação (n) é igual a 1,5 bp (espectroscopia de absorção), e n f ca. 2 bp (espectroscopia de fluorescência). Mostrou-se que n nf é dependente da quantidade de GC e n total é independente.The binding parameters of ethidium bromide (EtBr) with DNA of various GC/AT ratios were determined using absorption and fluorescence spectroscopy. Our experimental data clearly demonstrate the co-existence of fluorescing and non-fluorescing types of "strong" binding at low concentration of EtBr. The fluorescent complex corresponds to the ordinary intercalative model. The non-fluorescent complex is referred to semi-intercalative binding of EtBr. The binding constant (K) and the number of base pairs corresponding to a binding site (n) of the fluorescent (K f and n f ) and non-fluorescent (K nf , n nf ) types of interactions were determined. The average size of binding site (n) is equal to 1.5 bp (absorption spectroscopy), and n f ca. 2 bp (fluorescence spectroscopy). It was shown that n nf is dependent on GC-content and total n is independent of it.
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