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利用溴化乙锭作为荧光探针研究克百威与小牛胸腺 DNA 的相互作用的光谱研究。

Spectroscopic studies on the interaction between carbaryl and calf thymus DNA with the use of ethidium bromide as a fluorescence probe.

机构信息

State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China.

出版信息

J Photochem Photobiol B. 2012 Mar 1;108:53-61. doi: 10.1016/j.jphotobiol.2011.12.011. Epub 2012 Jan 4.

Abstract

The interaction between carbaryl and calf thymus DNA (ctDNA) was investigated under simulated physiological conditions (Tris-HCl buffer of pH 7.4) using ethidium bromide (EB) dye as a probe by UV-vis absorption, fluorescence and circular dichroism (CD) spectroscopy, as well as DNA melting studies and viscosity measurements. It can be concluded that carbaryl molecules could intercalate into the base pairs of DNA as evidenced by hyperchromic effect of absorption spectra, decreases in iodide fluorescence quenching effect, induced CD spectral changes, and significant increases in melting temperature and relative viscosity of DNA. The binding constants and thermodynamic parameters of carbaryl with DNA were obtained by the fluorescence quenching method. Furthermore, a chemometrics approach, parallel factor analysis (PARAFAC), was applied to resolve the measured three-way synchronous fluorescence spectral data matrix of the competitive interaction between carbaryl and EB with DNA, and the results provided simultaneously the concentration profiles and corresponding pure spectra for three reaction components (carbaryl, EB and DNA-EB complex) of the kinetic system at equilibrium. This PARAFAC analysis demonstrated the intercalation of carbaryl to the DNA helix by substituting for EB in the DNA-EB complex.

摘要

在模拟生理条件(pH 7.4 的 Tris-HCl 缓冲液)下,使用溴化乙锭(EB)染料作为探针,通过紫外可见吸收光谱、荧光和圆二色性(CD)光谱以及 DNA 热融研究和粘度测量,研究了carbaryl 与小牛胸腺 DNA(ctDNA)之间的相互作用。可以得出结论,carbaryl 分子可以嵌入 DNA 的碱基对中,这可以通过吸收光谱的增色效应、碘荧光猝灭效应的降低、诱导的 CD 光谱变化以及 DNA 熔点和相对粘度的显著增加来证明。通过荧光猝灭法获得了 carbaryl 与 DNA 的结合常数和热力学参数。此外,应用化学计量学方法平行因子分析(PARAFAC)来解析 carbaryl 和 EB 与 DNA 之间竞争相互作用的测量三通道同步荧光光谱数据矩阵,结果同时提供了动力学系统平衡时三个反应组分(carbaryl、EB 和 DNA-EB 复合物)的浓度分布和相应的纯光谱。这种 PARAFAC 分析证明了 carbaryl 通过取代 DNA-EB 复合物中的 EB 嵌入到 DNA 螺旋中。

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