本文選題：HIV + HIV-1　； 參考：《西北師范大學(xué)》2009年博士論文

【摘要】： SELEX技術(shù)是指數(shù)富集的配基系統(tǒng)進(jìn)化技術(shù)(systematic evolution of ligands by exponential enrichment,簡(jiǎn)稱SELEX技術(shù)),它是利用組合化學(xué)合成及篩選技術(shù),結(jié)合生物分子克隆分離技術(shù)的跨學(xué)科體外分子篩選技術(shù)體系。SELEX技術(shù)是突破傳統(tǒng)生物高分子必須來源于生物體的傳統(tǒng)制備研究方式。目前,利用該項(xiàng)技術(shù)已從核酸庫中篩選出能與多種靶分子特異結(jié)合的各種特異性配基,這類寡核苷酸配基具有廣泛的結(jié)合能力和卓越的特異性,成為生物高分子適配體研究的理想工具。 自從HIV(人類免疫缺陷病毒,亦稱艾滋病毒)二十世紀(jì)80年代被發(fā)現(xiàn)以來,由HIV引起的獲得性免疫缺陷病(AIDS)已成為一種嚴(yán)重威脅人類健康的重大疾病,造成2000多萬人的死亡。得益于現(xiàn)代分子生物學(xué)及相關(guān)學(xué)科先進(jìn)的研究手段,對(duì)于HIV的研究進(jìn)展神速,現(xiàn)已基本搞清了HIV基因結(jié)構(gòu)和病毒形態(tài),對(duì)其致病機(jī)理也有了深入的了解。由于HIV是一種逆轉(zhuǎn)錄病毒,它是一種全新的病毒類型,HIV的預(yù)防、診斷、治療至今仍是醫(yī)學(xué)領(lǐng)域的重大難題。如能采用非生物來源,通過化學(xué)合成的方法得到具有生物識(shí)別能力的智能高分子化合物,應(yīng)用于HIV的醫(yī)學(xué)和生物學(xué)研究,通過分子識(shí)別及靶向藥物產(chǎn)品的研究和開發(fā),對(duì)HIV的預(yù)防、診斷、治療具有非常重要的意義。 本課題的研究目的是采用組合化學(xué)SELEX技術(shù)篩選針對(duì)HIV-1病毒的p24抗原的特異性寡核苷酸配基生物高分子,并進(jìn)行化學(xué)修飾,體外活性鑒定及應(yīng)用等研究。 首先,利用組合化學(xué)SELEX技術(shù)對(duì)HIV-1病毒的p24抗原的特異核酸高分子配基進(jìn)行篩選;然后,采用分子生物學(xué)方法對(duì)核酸高分子配基進(jìn)行分離、分析及活性鑒定,得到高活性的核酸高分子配基;隨后,對(duì)核酸高分子配基進(jìn)行化學(xué)修飾及活性鑒定,完成在高分子配基上連接一個(gè)核酸信標(biāo)序列,使標(biāo)記后的配基成為具有配體識(shí)別和信號(hào)擴(kuò)增能力的智能檢測(cè)生物高分子。最后,完成了信標(biāo)配基智能生物高分子應(yīng)用研究。 綜上所述:本研究運(yùn)用組合化學(xué)SELEX技術(shù)首次篩選到19個(gè)HIV-1 p24抗原的特異性核酸高分子配基,首次構(gòu)建了HIV-1 p24抗原-核酸高分子信標(biāo)配基,并完成活性檢測(cè)及基礎(chǔ)應(yīng)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.
【學(xué)位授予單位】：西北師范大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2009
【分類號(hào)】：R373

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