有机小分子—核酸大分子相互作用研究及分析应用

发布时间：2018-02-27 04:11

本文关键词： 金纳米粒子适配体三磷酸腺苷氨基糖苷类抗生素 RRE RNA　出处：《陕西师范大学》2016年硕士论文　论文类型：学位论文

【摘要】：生物传感新方法的研究进展,推动了分析化学和生命医学的学科发展,在生命科学和分析化学的交叉领域,发挥着极为重要的作用。因此,大多数分析化学和生物医学的研究者致力于生物传感新方法的研究。核酸探针凭借着易于设计、信号转导灵活等优势受到了广泛的关注。本论文旨在基于核酸探针建立不同的传感方法。本论文包含以下三个部分：第一部分绪论首先对核酸探针的基本概念做了简单概述,尤其是核酸适配体及其基本特性和HIV RNA;其次对基于核酸探针各种分析方法进行了介绍；最后提出该论文的研究内容及目的。第二部分基于ATP-适配体复合物对金纳米粒子的保护作用比色检测三磷酸腺苷三磷酸腺苷(ATP)是生物体内最直接的能量来源,其检测在医药研究、生命科学、食品安全和环境监测等诸多领域都具有重要意义。本研究发现适配体-ATP复合物能够比单独的适配体DNA或ATP更好地保护金纳米颗粒以抵抗盐溶液引起的团聚。通过透射电镜、动态光散射、Zeta电位计和荧光偏振技术,该保护作用得到验证。以此为基础,提出了一种以ATP适配体作为识别元件,以未修饰的金纳米颗粒作为探针的高灵敏度比色传感检测ATP的方法。该传感方法在0.05μM至1μM的ATP浓度区间内,吸光度比值A680/A520与ATP浓度的对数表现出良好的线性关系。ATP类似物,如三磷酸鸟苷、三磷酸尿苷、三磷酸胞嘧啶对ATP的测定干扰较小或基本无干扰。第三部分基于小分子配体-HIV RNA相互作用检测氨基糖苷类抗生素小分子配体-功能性生物大分子相互作用的研究为发现有效调节特定生物过程的新化合物提供了机遇。自从发现核糖核酸(RNA)在调节基因表达方面的作用后,RNA便成为有吸引力的药物靶标。然而到目前为止,关于配体-功能性RNA相互作用的研究,尤其是基于荧光分子的研究是相对少见的,且氨基糖苷类抗生素的定量测定对临床医学和药物分析十分必要。本研究发现RRE RNA可有效猝灭普罗黄素的荧光,所以在研究中首先探讨了不同浓度RRE RNA对普罗黄素的猝灭及猝灭机理。有文献报道部分氨基糖苷类抗生素可以和HIV-1 RRE RNA特异性结合,进一步发现将氨基糖苷类抗生素加入到RRE RNA-普罗黄素的猝灭复合物中后,由于氨基糖苷类抗生素和RRE RNA相互作用导致普罗黄素被释放,使得普罗黄素的荧光得以恢复。因此,基于小分子和RRE RNA相互作用,以普罗黄素为荧光指示剂,建立了一种非标记检测氨基糖苷类抗生素的方法。
[Abstract]:The research progress of new biosensor methods has promoted the development of analytical chemistry and life medicine, and has played an extremely important role in the intersection of life science and analytical chemistry. Most analytical chemistry and biomedical researchers are working on new biosensors. Nucleic acid probes are designed with ease. The advantages of signal transduction flexibility have received wide attention. This paper aims to establish different sensing methods based on nucleic acid probes. This thesis consists of the following three parts: the first part is a brief introduction to the basic concepts of nucleic acid probes. In particular, the aptamers and their basic characteristics and HIV RNAs are introduced. Secondly, various analysis methods based on nucleic acid probes are introduced. In the second part, based on the protective effect of ATP-aptamer complexes on gold nanoparticles, colorimetric detection of adenosine triphosphate adenosine triphosphate (ATP) is the most direct energy source in vivo. Its detection in medical research, life sciences, In this study, it was found that aptamer -ATP complexes can better protect gold nanoparticles against salt solution agglomeration than individual aptamers DNA or ATP. The protective effect of Zeta potentiometer and fluorescence polarization technique is verified. Based on this, a ATP adaptor is proposed as a recognition element. The high sensitivity colorimetric sensing method using unmodified gold nanoparticles as probe was used to detect ATP. In the range of 0. 05 渭 M to 1 渭 M of ATP concentration, the absorbance ratio A680 / A520 showed a good linear relationship with the logarithm of ATP concentration. Such as guanosine triphosphate, uridine triphosphate, Cytosine triphosphate has little or no interference with the determination of ATP. Part 3: detection of small ligand ligand-functional biomolecules interaction of aminoglycoside antibiotics based on small ligand-RNA interaction. It offers an opportunity to identify new compounds that effectively regulate specific biological processes. Since the discovery of the role of RNA in regulating gene expression, RNA has become an attractive drug target. Studies on ligand-functional RNA interactions, especially those based on fluorescent molecules, are relatively rare. The quantitative determination of aminoglycoside antibiotics is necessary for clinical medicine and drug analysis. In this study, we found that RRE RNA can effectively quench the fluorescence of propranthin. Therefore, the quenching and quenching mechanism of profloxine at different concentrations of RRE RNA were studied. It was reported that some aminoglycoside antibiotics could bind to HIV-1 RRE RNA specifically. It was further found that when aminoglycoside antibiotics were added to the quenching complex of RRE RNA-profloxine, the fluorescence of profloxine was restored due to the interaction between aminoglycoside antibiotics and RRE RNA. Based on the interaction between small molecules and RRE RNA, a method for the detection of aminoglycoside antibiotics with profloxine as fluorescent indicator was developed.
【学位授予单位】：陕西师范大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：O641.3;O657.3

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