识别艾滋病毒（Ⅰ型）核心抗原p24的核酸高分子配基的制备、活性鉴定及应用

发布时间：2018-05-22 09:19

本文选题：HIV + HIV-1　；参考：《西北师范大学》2009年博士论文

【摘要】： SELEX技术是指数富集的配基系统进化技术(systematic evolution of ligands by exponential enrichment,简称SELEX技术),它是利用组合化学合成及筛选技术,结合生物分子克隆分离技术的跨学科体外分子筛选技术体系。SELEX技术是突破传统生物高分子必须来源于生物体的传统制备研究方式。目前,利用该项技术已从核酸库中筛选出能与多种靶分子特异结合的各种特异性配基,这类寡核苷酸配基具有广泛的结合能力和卓越的特异性,成为生物高分子适配体研究的理想工具。自从HIV(人类免疫缺陷病毒,亦称艾滋病毒)二十世纪80年代被发现以来,由HIV引起的获得性免疫缺陷病(AIDS)已成为一种严重威胁人类健康的重大疾病,造成2000多万人的死亡。得益于现代分子生物学及相关学科先进的研究手段,对于HIV的研究进展神速,现已基本搞清了HIV基因结构和病毒形态,对其致病机理也有了深入的了解。由于HIV是一种逆转录病毒,它是一种全新的病毒类型,HIV的预防、诊断、治疗至今仍是医学领域的重大难题。如能采用非生物来源,通过化学合成的方法得到具有生物识别能力的智能高分子化合物,应用于HIV的医学和生物学研究,通过分子识别及靶向药物产品的研究和开发,对HIV的预防、诊断、治疗具有非常重要的意义。本课题的研究目的是采用组合化学SELEX技术筛选针对HIV-1病毒的p24抗原的特异性寡核苷酸配基生物高分子,并进行化学修饰,体外活性鉴定及应用等研究。首先,利用组合化学SELEX技术对HIV-1病毒的p24抗原的特异核酸高分子配基进行筛选;然后,采用分子生物学方法对核酸高分子配基进行分离、分析及活性鉴定,得到高活性的核酸高分子配基;随后,对核酸高分子配基进行化学修饰及活性鉴定,完成在高分子配基上连接一个核酸信标序列,使标记后的配基成为具有配体识别和信号扩增能力的智能检测生物高分子。最后,完成了信标配基智能生物高分子应用研究。综上所述:本研究运用组合化学SELEX技术首次筛选到19个HIV-1 p24抗原的特异性核酸高分子配基,首次构建了HIV-1 p24抗原-核酸高分子信标配基,并完成活性检测及基础应用研究。
[Abstract]:SELEX is an exponentially enriched ligand phylogenetic technique called systematic evolution of ligands by exponential enrichment, for short SELEX. It is based on combinatorial chemical synthesis and screening techniques. The interdisciplinary molecular screening system, Selex, which combines with biological molecular cloning separation technology, is a traditional preparation method that breaks through the traditional biological polymer must be derived from the organism. At present, the technique has been used to screen various specific ligands from nucleic acid libraries that bind specifically to a variety of target molecules, and such oligonucleotide ligands have extensive binding ability and excellent specificity. It is an ideal tool for the study of biopolymer aptamers. Since the discovery of HIV (Human Immunodeficiency virus) in the 1980s, acquired Immunodeficiency Disease (HIV) has become a serious threat to human health, resulting in more than 20 million deaths. Thanks to the advanced research methods of modern molecular biology and related disciplines, the research on HIV has made rapid progress, and the structure of HIV gene and virus morphology have been basically understood, and the pathogenesis of HIV has also been deeply understood. Because HIV is a kind of retrovirus, it is a new type of virus, the prevention, diagnosis and treatment of HIV is still a major problem in the field of medicine. Intelligent polymer compounds with biometric ability can be obtained by chemical synthesis from non-biological sources. They can be used in the medical and biological research of HIV, and through the research and development of molecular recognition and targeted drug products. The prevention, diagnosis and treatment of HIV are of great significance. The purpose of this study was to select ligand oligonucleotide ligands against p24 antigen of HIV-1 virus by combined chemical SELEX technique, and to carry out chemical modification, in vitro activity identification and application. Firstly, the specific nucleic acid ligands of p24 antigen of HIV-1 virus were screened by combined chemical SELEX technique, and then the nucleic acid polymer ligands were isolated, analyzed and identified by molecular biology. The high activity nucleic acid polymer ligand was obtained, and then the nucleic acid polymer ligand was chemically modified and its activity was identified, and a nucleic acid beacon sequence was attached to the polymer ligand. The labeled ligands become intelligent detection biomolymers with the ability of ligand recognition and signal amplification. Finally, the application research of smart biomolymers with standard ligand was completed. To sum up: in this study, 19 specific nucleic acid ligands of HIV-1 p24 antigen were screened for the first time by combined chemical SELEX technique, and HIV-1 p24 antigen-nucleic acid macromolecule standard ligand was constructed for the first time, and the activity detection and basic application research were completed.
【学位授予单位】：西北师范大学
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R373

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