幽门螺杆菌中性白细胞激活蛋白基因克隆表达及免疫评价

发布时间：2018-04-02 01:28

本文选题：幽门螺杆菌　切入点：中性白细胞激活蛋白　出处：《郑州大学》2005年博士论文

【摘要】：幽门螺杆菌(Helicobacter pylori,H.pylori)是一种微需氧革兰氏阴性螺旋杆菌,在全球人口中的感染率超过50%。H.pylori感染多发生在儿童时期,如果不经治疗,H.pylori在胃中的持续慢性感染可导致胃炎、消化道溃疡等疾病,并与胃癌、胃黏膜相关性淋巴样组织(MALT)淋巴瘤发生密切相关。研制疫苗将是控制H.pylori感染及相关疾病经济有效的途径。 H.pylori水抽提物中存在着一种能趋化并激活中性白细胞的可溶性蛋白,被称作幽门螺杆菌中性白细胞激活蛋白(H.pylori neutrophil-activating protein,HP-NAP)。HP-NAP位于幽门螺杆菌细胞质中,可通过细菌自溶而释放,并附着在细菌外膜的表面,HP-NAP对高分子量碳水化合物具有特异亲和力,处于这样的位置HP-NAP可能介导幽门螺杆菌粘附于缩主胃黏膜上皮细胞表面。多数H.pylori感染者血清中存在HP-NAP特异抗体,HP-NAP被认为是幽门螺杆菌感染的重要因子和疫苗候选抗原。本研究利用PCR方法从H.pylori临床分离株MEL-HP27基因组DNA中克隆HP-NAP编码基因HP-napA,利用生物信息技术对HP-napA进行序列分析和功能预测,DNA重组技术构建原核表达载体,并对其编码产物的免疫原性、免疫反应性进行研究,从阻断H.pylori黏附与定植的角度观察HP-NAP在动物模型中的免疫保护效果,以评价其在发展H.pylori基因工程疫苗中的作用,为免疫接种预防H.pylori感染,减少H.pylori相关疾病奠定基础。实验方法 1.H.pylori MEL-HP27菌株HP-napA基因的分子克隆及序列分析
[Abstract]:Helicobacter pylori H.pylori, a microaerobic gram-negative helicobacter pylori, is more prevalent in the global population than in childhood, and a persistent chronic infection of H.pylori in the stomach without treatment can lead to gastritis. Gastrointestinal ulcer and other diseases are closely related to gastric cancer and gastric mucosa-associated lymphoid tissue lymphoma. The development of vaccine will be an economical and effective way to control H.pylori infection and related diseases. There is a soluble protein in H.pylori water extract that can chemoattractant and activate neutrophil. It is called H. pylori neutrophil-activating protein HP-NAPP. HP-NAP is located in the cytoplasm of Helicobacter pylori and can be released by bacterial autolysis. The HP-NAP attached to the bacterial outer membrane has specific affinity to high molecular weight carbohydrates. In this position, HP-NAP may mediate the adhesion of Helicobacter pylori to the surface of gastric epithelial cells. The presence of HP-NAP in the sera of most H.pylori infected patients is considered to be an important factor and vaccine candidate antigen for Helicobacter pylori infection. In this study, PCR method was used to clone the HP-NAP encoding gene HP-napA from the genomic DNA of H.pylori clinical isolate MEL-HP27. Sequence analysis and functional prediction of HP-napA were used to construct the prokaryotic expression vector, and the immunogenicity of the encoding product was analyzed. The immunoreactivity was studied to observe the immune protection effect of HP-NAP in animal model from the view of blocking the adhesion and colonization of H.pylori in order to evaluate its role in the development of H.pylori gene engineering vaccine and to prevent H.pylori infection. Reduce H.pylori related diseases to lay the foundation. Experimental method. Molecular cloning and sequence Analysis of HP-napA Gene of 1.H.pylori MEL-HP27 strain
【学位授予单位】：郑州大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R392

【参考文献】