新型核酸自组装纳米探针的构建及其在生物传感和成像分析中的应用

发布时间：2018-01-11 20:41

本文关键词：新型核酸自组装纳米探针的构建及其在生物传感和成像分析中的应用　出处：《青岛科技大学》2015年硕士论文　论文类型：学位论文

【摘要】：DNA纳米技术最早由纽约大学教授Seeman于1982年提出并展开研究,经过二十多年的发展,形成了DNA分子瓦自组装、折纸术和纳米排布三大研究方向,已成功制备出多种DNA纳米结构。核酸分子探针是由DNA和RNA构成的自组装纳米结构,凭借其易于标记、结构设计灵活、稳定等特点在生物传感中具有重要作用,可实现目标分子的识别、信号的转换和放大。一种多功能、可设计的DNA纳米结构可以引出多种新颖的核酸自组装路线,在生物传感、靶向给药、癌症治疗等应用领域有广阔的发展前景。本文针对核酸自组装技术开展了如下工作：一、核酸自组装纳米球的构建及在生物传感及成像分析中的应用本章通过碱基互补配对原则及DNA的液晶力,自组装得到粒径约为200 nm的单分散DNA纳米球结构。本方法较传统方法的优势在于,无需大量DNA链,只需四条DNA链即可完成纳米球的组装。纳米球中绝大多数的缺口可由T4连接酶连接,从而有效的催化了反应的进行,避免了核酸的分解,提高了DNA纳米球的生物稳定性。纳米球结构上可以组装多种功能基团,使其具有可设计性和固载量,可用于分子探针的构建、荧光成像分析、肿瘤细胞特异性识别、载药及药物传输、基因调节等,有望广泛应用于生物传感和肿瘤细胞的诊断与治疗。此外,该DNA纳米球还表现出良好的生物相容性,为DNA自组装、生物传感以及生物医学诊断和研究开辟了新的发展空间。二、基于四叉DNA纳米结构和DNA纳米球化学发光共振能量转移成像检测microRNA本章提出了一种准确检测microRNA的方法,利用microRNA引发发夹DNA构建DNA四叉结构,继而利用其自身的DNA酶序列实现核酸自组装,形成DNA纳米球,实现了对microRNA的高灵敏度、高特异性检测。从DNA纳米结构的构建出发,避免了microRNA碱基和反应条件的限制,通过DNA结构之间的互补,设计了多功能DNA发夹结构,通过DNA链取代反应将其组装成四叉DNA纳米结构,进而组装成DNA纳米球。在此过程中,microRNA可循环利用,从而实现microRNA的高效、特异性检测。
[Abstract]:DNA nanotechnology was first proposed and studied by New York University professor Seeman in 1982. After more than 20 years of development, DNA molecular tile self-assembly was formed. Origami and nanostructures have been successfully prepared. Nucleic acid probes are self-assembled nanostructures composed of DNA and RNA, which are easy to label. The characteristics of flexible structure design and stability play an important role in biosensor. It can realize the recognition of target molecules, signal conversion and amplification. The design of DNA nanostructures can lead to a variety of novel nucleic acid self-assembly routes in biosensor targeted drug delivery. Cancer therapy and other applications have a broad prospect. This paper focused on nucleic acid self-assembly technology carried out the following work: 1. Construction of nucleic acid self-assembled nanospheres and their applications in biosensor and imaging analysis this chapter uses the principle of base complementary pairing and the liquid crystal force of DNA. The monodisperse DNA nanospheres with a diameter of about 200nm were obtained by self-assembly. The advantage of this method over the traditional method is that there is no need for a large number of DNA chains. It takes only four DNA strands to complete the assembly of the nanospheres. Most of the gaps in the nanospheres can be connected by T4 ligase, which effectively catalyzes the reaction and avoids the decomposition of nucleic acids. The biological stability of DNA nanospheres is improved. The nanospheres can assemble a variety of functional groups in the structure, which can be used for molecular probe construction and fluorescence imaging analysis. Tumor cell specific recognition, drug delivery and drug delivery, gene regulation and so on, are expected to be widely used in the diagnosis and treatment of tumor cells. In addition, the DNA nanospheres also show good biocompatibility. For DNA self-assembly, biosensor and biomedical diagnosis and research opened up a new space for development. Based on Quadric DNA Nanostructures and DNA nanospheres chemiluminescence Resonance Energy transfer Imaging Detection microRNA this chapter presents an accurate detection method for microRNA. The quad structure of DNA was constructed by microRNA initiated hairpin DNA, and then the nucleic acid self-assembly was realized by using its own DNA enzyme sequence to form DNA nanospheres. The high sensitivity and specificity of microRNA detection was realized. Based on the construction of DNA nanostructures, the restriction of microRNA bases and reaction conditions was avoided. The multifunctional DNA hairpin structure was designed through the complementation of DNA structure, and the quad DNA nanostructure was assembled by the DNA chain substitution reaction, and then the DNA nanospheres were assembled. MicroRNA can be recycled to realize the high efficiency and specificity of microRNA detection.
【学位授予单位】：青岛科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TB383.1;TP212.3

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