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Electrochemical detection of abasic site-containing DNA

机译：电化学检测含碱基位点的DNA

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摘要

A simple and rapid approach for detecting apurinic (AP) sites in DNA, based on direct stripping chronopotentiometric measurements of the adenine and guanine nucleobases at a graphite electrode is described. Tetrahydrofuranyl residues, lacking a nucleobase moiety, were utilized for designing the AP sites and were incorporated in 19-mer oligonucleotides. The change of adenine-to-guanine response ratio (A/G) in one-. two- or three-substituted adenosine residues for stable analogs of AP sites was exploited for electrochemical measurements of the adenine loss. The resulting A/G response ratio decreases linearly upon increasing the number of AP sites in the oligonucleotides; the values of A/G electrochemical signals were slightly enhanced when compared to the actual purine content. HPLC analysis of the released nucleobases confirmed that the sulfuric acid-induced oliconucleotide cleavage provides complete apurination and dissolution of the released nucleobases in aqueous solution. Additional experiments with mixtures of free nucleobases and purine nucleosides reveal that the larger A/G ratio observed in the electrochemical analysis of AP-site-containing oligorners is attributed to the influence of the acid and/or thermal decomposition products (particularly the sugar fragments). This study represents the first step in developing a simple and direct electrochemical assay of AP sites in single-stranded DNA.
机译：描述了一种基于石墨电极上腺嘌呤和鸟嘌呤核苷碱基的直接剥离计时电位测量的简单快速的方法来检测DNA中的嘌呤（AP）位点。缺少核碱基部分的四氢呋喃基残基用于设计AP位点，并掺入19-mer寡核苷酸中。腺嘌呤与鸟嘌呤应答比（A / G）的变化为1。用于AP位点的稳定类似物的两个或三个取代的腺苷残基被用于腺嘌呤损失的电化学测量。随着寡核苷酸中AP位点数量的增加，所得的A / G响应比呈线性下降;与实际嘌呤含量相比，A / G电化学信号值略有提高。释放核苷的HPLC分析证实，硫酸诱导的寡核苷酸裂解可完全纯化和溶解释放的核碱在水溶液中。游离核碱基和嘌呤核苷混合物的其他实验表明，在含AP位的寡聚体的电化学分析中观察到的较大的A / G比归因于酸和/或热分解产物（特别是糖片段）的影响。这项研究代表着开发单链DNA中AP位点的简单直接电化学测定的第一步。

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