脑膜炎奈瑟氏菌表面蛋白A基因的克隆和表达

发布时间：2018-03-05 10:18

本文选题：脑膜炎奈瑟氏菌表面蛋白A　切入点：克隆和表达　出处：《中国医科大学》2006年硕士论文　论文类型：学位论文

【摘要】：脑膜炎奈瑟氏菌表面蛋白A基因的克隆和表达前言脑膜炎奈瑟氏菌是引起人流行性脑膜脑炎(以下简称流脑)的致病菌。在非流行期,发病率在1-3/10万左右,而在流行期,发病可高达500/10万,对6个月到2岁的婴幼儿及青少年危害极大,病死率高达10%以上,近年来,我国流脑发病率有明显上升趋势,暴发流行时有发生,对我国人民的生命健康构成极大威胁。脑膜炎奈瑟氏菌根据菌体表面的荚膜多糖分为12个血清群。全世界大约90%的病人是由A群、B群和C群引起的。以A群、C群荚膜多糖为基础的疫苗被开发出来并被证明对控制流脑的暴发和流行有效。但是A群、C群疫苗对幼儿缺乏免疫原性,而且它们提供的保护期相对短,一般为1-3年。B群荚膜多糖由于无法诱导人体产生免疫反应,所以B群流脑疫苗一直也没有被开发出来。基于这些原因,开发非荚膜多糖的表面蛋白抗原疫苗以刺激机体产生保护性、持久性抗体就成为另一种选择。但是每个血清群的脑膜炎奈瑟氏菌根据菌体主要表面蛋白(OMP)又可分为多个血清型、血清亚型,如B群有20多个血清型,每个血清型又有至少10多个血清亚型,依此类推12个血清群的脑膜炎奈瑟氏菌有上千种表面蛋白。寻找所有血清群共有的表面蛋白抗原成为几十年来全世界脑膜炎奈瑟氏菌研究者奋斗的目标,1997年Martin等发现了一种极为保守的表面蛋白,存在于所有的脑膜炎奈瑟氏菌菌体表面,命名为脑膜炎奈瑟氏菌表面蛋白A(NspA),含174个氨基酸,由525个核苷酸序列编码组成。尽管Ns-pA的功能至今还不清楚,但给小鼠注射纯化的或重组的NspA后都能诱导产生保护性抗体,抵抗脑膜炎奈瑟氏菌的攻击。综上所述,NspA对流脑的诊断和预防策略的制定具有极大的应用价值。为此,本研究对脑膜炎奈瑟氏菌表面蛋白A基因进行了克隆和表达,并采用ELISA对纯化的NspA融合蛋白进行了活性检测,同时验证了人体
[Abstract]:Cloning and expression of surface protein A gene of Neisseria meningitidis. Foreword. Neisseria meningitidis is the cause of human epidemic meningoencephalitis (hereinafter referred to as meningitis). It is harmful to infants and adolescents aged from 6 months to 2 years old, and the mortality is more than 10%. In recent years, the incidence of meningitis in China has an obvious upward trend, outbreaks and epidemics occur from time to time, which poses a great threat to the life and health of the people in our country. Neisseria meningitidis is divided into 12 serogroups based on capsular polysaccharides on the surface of the bacteria. About 90% of the world's patients are caused by groups A, B and C. Vaccine based on group A and group C capsule polysaccharides has been developed and developed. It has been proved to be effective in controlling the outbreak and epidemic of meningitis. But the group A and C vaccine is lack of immunogenicity in young children. And they offer relatively short protection periods, typically 1-3 years. Group B capsule polysaccharides have not been developed because they can't induce immune responses in humans. For these reasons, The development of a surface protein antigen vaccine for non-perfringent polysaccharides to stimulate protection in the body makes persistent antibodies an alternative. But each serotype of Neisseria meningitidis can be divided into several serotypes according to its main surface protein, serotypes, such as more than 20 serotypes in group B, and at least more than 10 serotypes in each serotype. And so on 12 serogroups of Neisseria meningitidis have thousands of surface proteins. Looking for surface protein antigens shared by all serums has been the goal of researchers around the world for decades, Martin et al., 1997. Found a very conservative surface protein, It exists on the surface of all Neisseria meningitidis. It is named as the surface protein of Neisseria meningitidis, AnspAN, which contains 174 amino acids and is composed of 525 nucleotide sequences, although the function of Ns-pA is still unknown. But injected with purified or recombinant NspA could induce the production of protective antibodies against Neisseria meningitidis. To sum up, NspA has great application value in the diagnosis and prevention of meningitis. Therefore, the surface protein A gene of Neisseria meningitidis was cloned and expressed in this study. The activity of purified NspA fusion protein was detected by ELISA, and the human body was verified.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R378

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