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一种研究多环芳烃在DNA溶液中增溶作用的简单方法。

A simple method for studying the solubilization of polycyclic aromatic hydrocarbons in DNA solutions.

作者信息

Chen F M

出版信息

Anal Biochem. 1983 Apr 15;130(2):346-52. doi: 10.1016/0003-2697(83)90598-5.

Abstract

A simple method for detecting the amount of a polycyclic aromatic hydrocarbon (PAH) solubilized in DNA solutions is described. This method takes advantage of the fact that sodium dodecyl sulfate (SDS) induces dissociation of PAH from DNA, and the greatly enhanced solubility of PAH in SDS solution makes possible the direct preparation of a standard solution for the extinction coefficient determination. The ability to make direct comparison between the spectroscopic characteristics before and after dissociation is accomplished by utilizing a tandem cuvette. The usefulness of this technique is then demonstrated by applying it to solubilization studies of pyrene in native and denatured DNA solutions of varying sodium chloride concentrations as well as in deoxymononucleotide solutions. Binding constants are estimated and the solubility data for pyrene in the DNA solutions are interpreted in terms of the binding abilities of intercalative versus external sites and the effect of salt. The mononucleotide results suggest a binding preference of pyrene to purine bases. It was observed that the binding constant of mononucleotide to pyrene is an order of magnitude lower than that of single-stranded DNA, which in turn is an order of magnitude smaller than that of duplex DNA.

摘要

本文描述了一种检测溶解于DNA溶液中的多环芳烃(PAH)含量的简单方法。该方法利用了十二烷基硫酸钠(SDS)能诱导PAH从DNA上解离这一事实,且PAH在SDS溶液中的溶解度大幅提高,使得直接制备用于测定消光系数的标准溶液成为可能。通过使用串联比色皿,可以直接比较解离前后的光谱特征。然后将该技术应用于不同氯化钠浓度的天然和变性DNA溶液以及脱氧单核苷酸溶液中芘的增溶研究,证明了该技术的实用性。估算了结合常数,并根据嵌入位点与外部位点的结合能力以及盐的影响,对芘在DNA溶液中的溶解度数据进行了解释。单核苷酸的结果表明芘对嘌呤碱基有结合偏好。观察到单核苷酸与芘的结合常数比单链DNA低一个数量级,而单链DNA又比双链DNA小一个数量级。

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