赖型钩端螺旋体毒力相关基因ompl17的蛋白质功能学研究

发布时间：2018-04-09 10:32

本文选题：钩端螺旋体　切入点：毒力相关基因　出处：《四川大学》2006年博士论文

【摘要】：钩端螺旋体病(简称钩体病)是由钩端螺旋体引起的一种人兽共患自然免疫源性疾病，它在世界范围内流行，并严重危害人类的健康和农牧业生产。钩端螺旋体(简称钩体)在结构和遗传特征上都比较特殊，因而大多数细菌学研究方法不适于钩体研究。随着中国南方中心对赖型钩体56601的测序完成，为钩体病的研究提供了新的契机，但是与钩体致病或毒力相关的因子研究报道还相对较少，，致使钩体致病的分子机制研究进展缓慢。因此，阐明钩端螺旋体的致病机制是控制本病的关键环节。本室曾以抑制消减杂交技术(Substration Suppressive Hybridization，SSH)筛选得到致病钩体与非致病钩体的差异基因片段，并采用盒式连接半巢式PCR技术扩增出ompl17基因，进行序列测定、生物信息学分析，同时采用基因打靶技术进行了功能的初步分析和探讨。本研究既往工作的基础上，对赖型钩体ompl17基因进行扩增表达，制备抗血清，分析该基因在赖型钩体体内外和不同温度下表达情况，同时对该蛋白进行表达纯化，采用构建血管内皮细胞基底膜进行血细胞的趋化分析，以及采用酵母双杂交技术钓取ompl17基因在体内相互作用的靶蛋白，为阐明钩体分子致病机制作出新的探索。
[Abstract]:Leptospirosis (referred to as leptospirosis) is a zoonotic disease caused by natural immunogenicity of Leptospira, it is popular in the world, and serious harm to human health and animal husbandry. Leptospira (leptopira) on the structure and genetic characteristics are quite special, so most bacteriological study method is not suitable for Leptospira research. With China Center of Leptospira Lai South 56601 sequencing, provides a new opportunity for the study of leptospirosis, but factor related research reports and Leptospira virulence is relatively less, the research progress in molecular mechanisms of the pathogenic Leptospira slowly. Therefore, to clarify the pathogenic mechanism of Leptospira the body is the key to control the disease.
The room was by suppression subtractive hybridization (Substration Suppressive Hybridization, SSH) were screened between pathogenic and non pathogenic Leptospira gene fragment, and connected with a semi nested PCR ompL17 gene was amplified by cassette, sequenced, bioinformatics analysis, and the function of the preliminary analysis and discussion by gene targeting. Based on the previous work, the Lai Leptospira ompL17 gene amplified expression, antiserum preparation, analysis and expression of the gene in Leptospira Lai in vivo at different temperatures and at the same time, the protein was expressed and purified, analyzed by chemotactic blood cells to construct vascular endothelial cells of the basilar membrane, and the use of the yeast two hybrid technique to catch the target protein ompL17 gene in vivo interaction, make new exploration in order to elucidate the molecular pathogenesis of Leptospira.

【学位授予单位】：四川大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R377

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