HMGB1结构域和HMGB1A盒与LBPK95A融合基因的克隆表达及其生物学功能的初步研究

发布时间：2018-01-30 21:03

本文关键词： 高迁移率族蛋白1A盒/B盒 LBPK95A 原核表达人单核细胞肿瘤坏死因子α 脓毒症　出处：《第四军医大学》2005年硕士论文　论文类型：学位论文

【摘要】：脂多糖(LPS)是革兰阴性菌(GNB)细胞壁的主要成份,是GNB所致脓毒症多种病理生理反应的第一触发因子。其引起的脓毒症和脓毒性休克是严重危及生命的全身性炎性反应综合征,致死率在儿童及成年人中分别达10～15%和40%左右。探讨脓毒症的新的有效防治途径无疑具有重要意义。近年研究发现,高迁移率族蛋白1(HMGB1)作为脓毒症潜在的晚期炎症介质参与了内毒素的致病过程,是脓毒症致死效应的重要炎症介质,对HMGB1干预可有效阻止脓毒症。HMGB1结构域B盒在炎症过程中的生物学功能与全长HMGB1相似,在HMGB1的致炎作用中发挥巨大作用。HMGB1结构域A盒具有类似HMGB1抗体的作用,可抑制细胞外HMGB1的活性,减少TNF-α等炎症介质的释放,控制脓毒症的发生发展。LPS结合蛋白(LBP)作为LPS的转运蛋白,控制着LPS诱导的单核/巨噬细胞炎性反应。LBP结构域LBPK95A可阻断LPS与LBP的结合,抑制LPS的细胞毒作用。如果将HMGB1 A盒和LBPK95A两者结合起来,从脓毒症的级联反应的早期和晚期进行干预,有可能为脓毒症的防治提供新的有效途径。为此本课题进行了以下主要研究:鼠高迁移率族蛋白1结构域A盒、B盒编码序列的克隆和原核表达及生物学活性鉴定;HMGB1A盒与LBPK95A融合基因的表达及纯化;A-LBP融合蛋白生物学功能的初步研究。一、HMGB1 A盒及B盒编码序列的克隆表达、产物纯化和活性鉴定
[Abstract]:Lipopolysaccharide (LPS) is the main component of the cell wall of Gram-negative bacteria (GNB). Sepsis and septic shock are the first trigger factors of pathophysiological responses to sepsis induced by GNB, which is a serious life-threatening systemic inflammatory response syndrome. The mortality rate in children and adults is about 1015% and 40% respectively. It is of great significance to explore the new effective prevention and treatment of sepsis. High mobility group protein (HMGB1), as a potential late inflammatory mediator of sepsis, plays an important role in the pathogenesis of endotoxin and plays an important role in the lethal effect of sepsis. Intervention with HMGB1 could effectively prevent the biological function of sepsis. HMGB1 domain B box in inflammatory process similar to that of full-length HMGB1. HMGB1 domain A box has the similar effect of HMGB1 antibody and can inhibit the activity of extracellular HMGB1. Reduce the release of inflammatory mediators such as TNF- 伪, and control the occurrence and development of sepsis. LPS-binding protein (LBP) is the transporter of LPS. Control of monocyte / macrophage inflammatory response induced by LPS. LBP domain LBPK95A could block the binding of LPS to LBP. Inhibits the cytotoxicity of LPS. If you combine HMGB1 A box with LBPK95A, you can intervene in the early and late stages of the cascade of sepsis. It is possible to provide a new effective way for the prevention and treatment of sepsis. Therefore, this paper has carried out the following main research: mouse high mobility group protein 1 domain A box. Cloning, prokaryotic expression and biological activity identification of B box coding sequence; Expression and purification of HMGB1A cassette and LBPK95A fusion gene; Preliminary study on biological function of A-LBP fusion protein. Cloning, expression, purification and activity Identification of coding sequences of HMGB1 A Box and B Box
【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R346

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