【作者】 尚智美；

【导师】 孙伟；

【作者基本信息】 青岛科技大学 ， 分析化学， 2006， 硕士

【摘要】 本论文用巯基自组装法制备了DNA电化学传感器，分别以亚甲基蓝和甲苯胺蓝为杂交指示剂，用紫外可见光谱和电化学法研究了它们与单双链DNA相互作用的识别模式，建立了电化学生物传感器法测定转基因NOS序列的新方法，还利用线性扫描极谱法研究了甲基紫与DNA的相互作用。 论文主要由五部分组成，第一部分综述了转基因植物产品、DNA电化学传感器的研究方法和DNA与小分子化合物的作用及意义。第二部分利用循环伏安法初步研究了巯基乙酸自组装膜修饰金电极的电化学行为，结果表明巯基乙酸在金电极表面具有特性吸附，形成具有一定致密性的自组装层。第三部分制备了巯基乙酸自组装膜DNA电化学传感器，以亚甲基蓝为杂交指示剂，用循环伏安法检测杂交前后其峰电流的变化，并应用于NOS基因片段的检测。第四部分详细研究了甲苯胺蓝在单双链DNA修饰电极上的电化学行为，并以甲苯胺蓝为杂交指示剂，利用制得的DNA电化学传感器对转基因NOS序列进行了测定。第五部分用线性扫描极谱法研究了甲基紫与DNA的相互作用，优化了结合反应条件和电化学测定条件，探讨了结合反应的机理，建立了以甲基紫为电化学探针极谱法测定DNA的新方法，并将该方法用于合成样品中DNA的检测，结果令人满意。更多还原

【Abstract】 In this paper, a gene electrochemical biosensor was prepared by self-assembled method, using the methylene blue (MB) and toluidine blue (TB) as the indicator respectively. The interaction model of indicator with ssDNA or dsDNA was investigated by UV-Vis adsorption spectrophotometry and cyclic voltammetry. The interaction of methyl violet (MV) with fsDNA was also investigated by linear sweep voltammetric technique.The paper is made up of five parts. Firstly, a summary referring to the genetically modified organism, the significance of the electrochemical biosensor, and the interaction of DNA with small molecules were reviewed. Secondly, the electrochemical behaviors of self-assembled modified gold electrode with mercaptoacetic acid were preliminarily studied by cyclic voltammetric technique. The results showed that a self-assembled monolayer was formed on the surface of gold electrode. Thirdly, a NOS gene electrochemical biosensor was prepared by covalently immobilizing of single-stranded NOS ssDNA sequence on the mercaptoacetic acid monolayer self-assembled gold electrode with the help of N-(3-dimethyIaminopropyl) -N’-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). The electroactive indicator, methylene blue (MB), is used to monitor the recognition of the DNA. Forthly, the interaction model of toluidine blue (TB) with ssDNA or dsDNA was更多还原