问号状赖型钩端螺旋体OmpA膜蛋白Loa22基因克隆及表达研究

发布时间：2018-04-30 03:38

本文选题：钩端螺旋体 + OmpA基因　；参考：《四川大学》2006年硕士论文

【摘要】：钩端螺旋体病(简称钩体病)是在世界范围内流行的人兽共患自然疫源性疾病。我国是钩体病疫情较为严重的国家，钩体病的防治是一项重要的任务。钩端螺旋体病的预防目前以疫苗为主，但现有疫苗仅能产生短期的、不完全的免疫保护效应，因此对赖型钩端螺旋体(简称钩体)新型疫苗进行研究，有着重要的现实意义。细菌的外膜成分由于位置的重要性往往成为疫苗研制的首要选择。OmpA是一种在肠科杆菌中高度保守的外膜蛋白，它由三个功能性的结构域组成：一个亲水的胞外结构域，一个跨膜的β桶状结构域，一个肽聚糖结构域。OmpA能在无免疫佐剂辅助的情况下诱导特异性的体液和细胞毒性反应，而且还能辅助其它抗原(如Ovalbumin，卵白蛋白)内化和交叉递呈，具有潜在的免疫载体功能，能用于疫苗的研究。我们以赖型钩体中编码OmpA膜蛋白的Loa22基因为对象，对其进行克隆分析，并对其进行原核及真核表达。在本研究中，，我们首先以问号状赖型钩体017株、56601株及双曲钩体Patoc Ⅰ株基因组为模板PCR扩增目的基因。随后在原核表达方面，我们构建了Loa22基因与质粒pGEX-4T-1的原核重组表达质粒
[Abstract]:Leptospirosis (Leptospirosis) is a worldwide epidemic of zoonotic diseases. Leptospirosis is a serious epidemic situation in China, and the prevention and control of leptospirosis is an important task. The main prevention of leptospirosis is vaccine, but the existing vaccine can only produce short-term and incomplete immune protection effect. Therefore, it is of great practical significance to study the new vaccine of Leptospira Leptospira (Leptospira Leptospira). The outer membrane components of bacteria are often the primary choice for vaccine development because of the importance of their location. OmpA is a highly conserved outer membrane protein in Enterobacteriaceae, which consists of three functional domains: a hydrophilic extracellular domain. A transmembrane 尾 barrel domain, a peptidoglycan domain. OmpA can induce specific humoral and cytotoxic responses without immune adjuvant, but also assist in the internalization and cross-presentation of other antigens, such as Ovalbumin (ovalbumin). It has potential immune vector function and can be used in vaccine research. The Loa22 gene encoding OmpA membrane protein in Leptospira lanceolaris was cloned and analyzed for prokaryotic and eukaryotic expression. In this study, we first amplified the target gene by PCR from the genomes of 56601 strain of Leptospira interrogans 017 and Patoc I strain of hyperbolic Leptospira. Then, in prokaryotic expression, we constructed the prokaryotic expression plasmid of Loa22 gene and plasmid pGEX-4T-1.
【学位授予单位】：四川大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R377

【引证文献】