【作者】 杨涛；

【导师】 焦奎；

【作者基本信息】 中国海洋大学 ， 海洋化学， 2006， 博士

【摘要】 近几年来,DNA电化学生物传感器作为一种新的DNA生物传感器,在检测特定序列的DNA时以其快速、简单、廉价而备受关注,发展十分迅速。根据电化学DNA传感器的制作目的,大致可以分为两类:一类是电化学DNA杂交传感器也可以称为电化学基因识别传感器,它是在电极表面固定单链DNA(ssDNA)作为探针,实现对与探针互补的目标DNA的检测。另一类则是非基因识别的电化学DNA传感器,它是在电极表面固定双链DNA(dsDNA)作为传感器的敏感器件,它的作用在于其它物质与敏感元件的作用或者利用DNA的特性来实现对特定物质的检测和研究,如研究DNA的结构、小分子与DNA的相互作用、金属离子的测定以及电催化应用等。这其中电化学基因识别传感器是当前研究的热点。制备电化学基因识别传感器,首先必须将DNA探针固定到电极表面,然后通过DNA杂交检测探针的互补序列。目前已经发展起来了各种各样的DNA固定化技术,如表面吸附法,共价键合法,自组装法,电聚合法和组合法等。其中自组装法和电聚合法可以综合利用共价键合法和吸附法将DNA固定于电极上。本论文制备了三种聚合物膜和一种自组装膜修饰电极用于固定DNA。论文分为三个部分,分别为前言部分、第一部分(三种聚合物膜制备DNA电化学传感器的研究)、第二部分(基于对-巯基苯胺自组装膜的DNA电化学传感器的构建及其应用)。前言部分首先系统介绍了DNA的结构及DNA的分子导电性。其次介绍了DNA电化学生物传感器的研究进展和分类,DNA电化学传感器的设计(DNA片段的固定方法以及杂交信号的转化两方面)以及在基因检测方面的应用和今后的发展趋势。其中在固定方法方面着重介绍了自组装法和电聚合法,在信号转换方面着重介绍了无指示剂型基于交流阻抗技术的构建的电化学DNA生物传感器。随后介绍了非基因识别型DNA电化学传感器在对DNA结构的研究、环境监测、金属离子的测定、药物的检测、筛选及抗病机理的研究等方面的应用。最后综合文献提出目前聚合物膜与自组装膜修饰电极用于固定DNA所存在的问题以及本课题的创新更多还原

【Abstract】 DNA electrochemical sensors hold an enormous potential for disease diagnosis, drug screening or forensic applications and development of an inexpensive, easy-to-use, fast response device stays therefore in the center of interest of many scientists. At present, many new electrochemical procedures are under intense investigation in order to get a fast, sensitive and selective quantification of nucleic acids and to meet the new challenges connected to the application of DNA biosensors in different fields. Among these new methods, electrochemical sensors offer powerful tools for converting the hybridization event into an analytical signal. They have been proposed in many reports for detecting the DNA hybridization event, due to their high sensitivity, small dimensions, low cost, and compatibility with microfabrication technology.The immobilization of ssDNA at the electrode is the key step in the preparation of DNA biosensors. Many methods have been employed for the immobilization of DNA, including adsorption, self-assembly monolayer (SAM), covalent binding, blending and electropolymerization. Self-assembly monolayer and electropolymerization synthesize adsorption and covalent binding method. Immobilization of a single-stranded (ss) deoxyribonucleic acids (DNA) chain or an oligonucleotide of specific sequence on the electrode creates a biosensor that detects the DNA hybridization. Another type of device is a biosensor with immobilized double-stranded (ds) DNA detecting small molecules interacting with DNA. This kind of biosensor is supposed to be used for detecting harmful substances present, e.g., in the environment as well as in food and so on.The goal of the present study is to design and optimize new DNA immobilization更多还原