甲型副伤寒沙门氏菌外膜蛋白的表达及其免疫保护性研究

发布时间：2018-01-12 08:13

本文关键词：甲型副伤寒沙门氏菌外膜蛋白的表达及其免疫保护性研究　出处：《南昌大学》2007年硕士论文　论文类型：学位论文

【摘要】： 伤寒是一种急性全身性感染，每年全球伤寒(包括副伤寒)患者约2200万，死亡超过20万人。伤寒包括伤寒沙门氏菌(Salmonella typhi)引起的伤寒和甲、乙、丙型副伤寒沙门氏菌(Salmonella paratyphi A，，B and C)引起的副伤寒。近年来我国甲型副伤寒的发病率呈逐年上升趋势，研究出安全、高效、经济的疫苗，对于预防甲型副伤寒爆发具有重大的经济社会效益。甲型副伤寒沙门氏菌是无荚膜的革兰氏阴性菌，其外膜蛋白在致病和刺激机体免疫应答方面起着非常重要的作用。因此，在研究和开发疫苗中，病原体的外膜蛋白成为热点。本论文对甲型副伤寒沙门氏菌外膜蛋白的克隆表达及其免疫保护性进行了研究，目的在于探索应用甲型副伤寒沙门氏菌外膜蛋白作为保护性抗原，获得甲型副伤寒候选疫苗的可能性。本课题选择毒力较强的甲型副伤寒沙门氏菌50973做为实验菌株，以NmpC、MipA、OmpW、PagC、OmpX、PomP(GI：56412276)、OmpC、FadL、TolC、BtuB和Imp等11个外膜蛋白为目标蛋白，分别将其基因的PCR扩增片段克隆到pET-32a表达载体，筛选出阳性克隆，用IPTG作为诱导剂，实现了这11种目标外膜蛋白的过量表达。利用这些过表达目标蛋白在盐溶液中形成沉淀而在水中可溶的特性实现了目标蛋白的有效分离。采用腹腔注射和体内攻毒的方法对纯化好的外膜蛋白的免疫保护性进行了研究，结果表明，PagC、OmpX、NmpC、FadL、TolC和BtuB等6个外膜蛋白具有良好的免疫保护力，其保护率分别为100％、100％、95％、75％、75％和70％。
[Abstract]:Typhoid is an acute systemic infection that affects about 22 million people worldwide each year with typhoid fever (including paratyphoid fever). More than 200,000 people were killed. Typhoid includes typhoid fever caused by Salmonella typhimurium (Salmonella typhimurium). Paratyphoid fever caused by Salmonella paratyphi Agna B and (C). In recent years, the incidence of paratyphoid A in China has been increasing year by year. The study of a safe, efficient and economical vaccine has great economic and social benefits for the prevention of paratyphoid A (paratyphoid A) outbreak. Salmonella paratyphoid A is a gram-negative bacteria without capsule, and its outer membrane protein plays a very important role in pathogenic and stimulating immune response. Therefore, in the research and development of vaccines. The outer membrane protein of pathogens has become a hot spot. In this paper, the cloning and expression of the outer membrane protein of Salmonella paratyphi A and its immune protection were studied. The purpose of this study was to explore the application of the outer membrane protein of Salmonella paratyphoid A as a protective antigen. The possibility of obtaining a candidate vaccine for paratyphoid A (paratyphoid A). In this study, the virulent Salmonella paratyphoid A (50973) was selected as the experimental strain, and NmpCT-MipAmpWPagCor OmpX was used as the experimental strain. Pom Pi: 56412276OmpCy FadLn TolCtuB and Imp were the target proteins. The PCR fragments of the gene were cloned into the pET-32a expression vector, and the positive clones were screened out. IPTG was used as the inducer. The overexpression of the 11 target outer membrane proteins was realized. The effective separation of the target proteins was achieved by intraperitoneal injection and in vivo, using the characteristics of the over expressed proteins precipitating in salt solution and soluble in water. The immunological protection of purified outer membrane protein was studied by the method of virus attack. The results showed that the six outer membrane proteins, such as PagCon OmpX BtuB, Fad LT TolC and BtuB, had good immunological protection, and the protective rates were 100% and 100%, respectively. There are 75 percent and 70 percent.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R378

【引证文献】