诱导型一氧化氮合酶基因调节蛋白的筛选及调节蛋白XBP1功能和启动子活性研究

发布时间：2018-02-13 07:43

本文关键词： 诱导型一氧化氮合酶 X-box结合蛋白1 CAT ELISA 转录调控启动子　出处：《重庆医科大学》2006年博士论文　论文类型：学位论文

【摘要】： 研究目的真核细胞中一氧化氮(NO)作为第二信使和神经递质发挥着多种不同的功能,具有广泛的生物学效应和抗病毒作用,参与机体的多种生理及病理过程,其对疾病发生及发展的影响是近年来研究的热点。诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)是产生一氧化氮的限速酶,广泛分布于机体内,一切影响iNOS活性的因素及影响iNOS基因转录、翻译的因素都会影响一氧化氮的生成,其中iNOS基因转录过程的调控是一氧化氮产量控制的主要因素。本研究首先运用噬菌体表面展示技术筛选出12种iNOS基因的调节蛋白,X-盒结合蛋白1(X-box binding protein1,XBP1)是其中一种iNOS基因的调节蛋白,他是CREB/ATF家族一个重要的转录因子,通过与真核基因启动子区域中顺式作用元件发生特异性相互作用、或者与其他相关蛋白质之间发生相互作用,激活或抑制转录,调节基因的表达,并通过整合这些不同的调节机制,激发无数基因的表达模式。为揭示真核细胞中iNOS基因的转录调控机制、反式作用因子XBP1对iNOS基因的调节作用,我们进一步针对筛选出的调节蛋白XBP1进行生物学功能的初步研究,对XBP1进行细胞内定位;筛选和构建XBP1与其他蛋白质间相互作用的连锁图,为阐明XBP1基因和iNOS基因的转录调节机制提供实验依据,为进一步揭示NO在肝细胞中的信号传递机制奠定理论基础。研究方法
[Abstract]:Research purpose. Nitric oxide (no) in eukaryotic cells, as a second messenger and neurotransmitter, plays many different functions, has a wide range of biological and antiviral effects, and participates in various physiological and pathological processes of the body. Inducible nitric oxide synthase (inducible nitric oxide synthase iNOS) is a rate-limiting enzyme that produces nitric oxide and is widely distributed in the body. All factors that affect the activity of iNOS and the transcription of iNOS gene are involved in this study. Translation factors affect the production of nitric oxide, The regulation of the transcription of iNOS gene is the main factor to control the production of nitric oxide. In this study, 12 iNOS gene regulatory proteins (1 X box binding protein 1 X box binding protein 1 XBP1) were screened by phage display technique. The regulatory protein of the iNOS gene, He is an important transcription factor in the CREB/ATF family that activates or inhibits transcription by interacting specifically with cis-acting elements in the eukaryotic promoter region or with other related proteins. In order to reveal the transcriptional regulation mechanism of iNOS gene in eukaryotic cells, the transactional factor XBP1 regulates the expression of iNOS gene by integrating these different regulatory mechanisms. We further studied the biological function of the screened regulatory protein XBP1, and carried out intracellular localization of XBP1. We screened and constructed a linkage map of the interaction between XBP1 and other proteins. In order to elucidate the transcriptional regulation mechanism of XBP1 gene and iNOS gene, and lay a theoretical foundation for further revealing the signal transduction mechanism of no in hepatocytes. Research method
【学位授予单位】：重庆医科大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R346

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