幽门螺杆菌ureB基因及其片段的表达和免疫研究

发布时间：2018-11-16 17:50

【摘要】：幽门螺杆菌(Helicobacter pylori，Hp)是一种革兰氏阴性菌、呈螺旋形，是引起人发生慢性活动性胃炎、胃和十二指肠溃疡的主要病原菌，同时与胃腺癌、胃粘膜相关淋巴组织淋巴瘤(MALT)的发生也密切相关。临床现有的根除Hp的手段主要是联合应用抗生素，但由于该菌感染的普遍性、耐药菌株的不断增加、较高的花费以及病人的低依从性等，限制了抗菌药物的疗效，因此，迫切需要进行免疫防治研究工作。幽门螺杆菌尿素酶是一种高亲和力、高活性和高分子质量的蛋白质。纯化尿素酶是由尿素酶A亚单位和B亚单位组成。尿素酶具有保护力、免疫原性、表面暴露和高度保守性，是幽门螺杆菌的主要抗原成分，，尤其是尿素酶B亚单位(UreB)，已经被证明可以刺激机体产生免疫应答且具有较好的免疫保护性。但是，由于UreB分子较大，在大肠杆菌中不易重组表达，而且UreB的重组表达产物多为包含体形式，表达蛋白失去活性而且提取工艺复杂。因此，我们以本实验室构建的具有分泌表达系统的大肠杆菌基因工程菌，即转入ureB及其e、f、m三个不同片段的大肠杆菌为研究对象，分析不同重组蛋白(rUreB、E、F、M)的表达和免疫性质。 1、借助生物信息学技术对重组蛋白理化性质、二级、三级结构
[Abstract]:Helicobacter pylori (Helicobacter pylori,Hp) is a gram-negative, spirally shaped bacterium that causes chronic active gastritis, gastric and duodenal ulcers, as well as gastric adenocarcinoma. The occurrence of gastric mucosa-associated lymphoid tissue lymphoma (MALT) is also closely related. The main method to eradicate Hp in clinic is the combination of antibiotics. However, the prevalence of infection, the increasing of drug-resistant strains, the high cost and the low compliance of patients limit the curative effect of antimicrobial agents. There is an urgent need for research on immunization prevention and control. Helicobacter pylori urease is a kind of protein with high affinity, high activity and high molecular weight. The purified urease consists of subunit A and subunit B of urease. Urease is protective, immunogenicity, surface exposure and highly conserved. It is the main antigen of Helicobacter pylori, especially the urease B subunit (UreB),. It has been proved that it can stimulate the body to produce immune response and has better immune protection. However, because of the large molecular size of UreB, it is difficult to express in E. coli, and the recombinant expression products of UreB are mostly in the form of inclusions, so the expressed protein is inactive and the extraction process is complex. Therefore, we studied the recombinant Escherichia coli strain constructed in our laboratory with secretory expression system, that is, E. coli, which was transferred into ureB and three different fragments of Erythrofon m, and analyzed different recombinant proteins (rUreB,E,F,). M) expression and immunological properties. 1. The physicochemical properties, secondary and tertiary structures of recombinant proteins were studied by bioinformatics.
【学位授予单位】：浙江工商大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R378;Q78

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