黑龙江立克次体外膜蛋白B基因表达及表达重组蛋白的抗原性分析  

Antigenicity Analysis of Recombinant Outer Membrane Protein B of Rickettsia heilongjiangensis

作　　者：孟艳芬[1] 段长松[1] 王锡乐[1] 熊小路[1] 温博海[1] 

机构地区：[1]军事医学科学院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京100071

出　　处：《寄生虫与医学昆虫学报》2011年第1期28-33,共6页Acta Parasitologica et Medica Entomologica Sinica

基　　金：国家基础研究计划项目（973计划课题 2010CB530200/2010CB530205）;国家科技重大专项（重大传染病预防与控制课题2008ZX10004-002）

摘　　要：黑龙江立克次体为远东蜱传斑点热病原体,本文以黑龙江立克次体基因组为模板,扩增ompB的4段基因,利用原核表达载体构建重组表达质粒,IPTG诱导重组质粒转化的大肠杆菌表达重组目的蛋白,并对其抗原性进行分析.免疫印迹反应表明,重组表达的4个蛋白均与黑龙江立克次体感染小鼠血清反应.该4个重组蛋白分别免疫小鼠后,分析其血清的特异性结果显示,4个重组蛋白免疫血清,其特异性抗体效价均≥1：5 120,与斑点热群立氏立克次体有相对明显的血清学交叉反应（≥1：40）,而和斑疹伤寒群立克次体仅有轻度交叉反应（≤1：40）.结果表明,重组OmpB蛋白能有效诱导机体产生特异性体液免疫应答,并能与黑龙江立克次体抗体产生特异反应,显示其具有良好的抗原性.Rickettsia heilongjiangensis is a human pathogen that causes Far East tick-borne spotted fever. Four fragments of outer membrane protein B gene （ompB） were amplified from R. heilongjiangensis, and the gene fragments were recombined with prokaryotic expression plasmid. Four recombinant OmpB proteins were expressed in E. coli cells transformed with the recombinant plasmids, respectively, following IPTG stimulation. In immunoblot assay, the recombinant OmpB proteins reacted with sera from mice infected with R. heilongfiangensis. In immunofluorescence assay, the sera from mice immunized with each recombinant OmpB protein strongly reacted with R. heilongjiangensis and the specific antibody titers were≥ 1：5 120, and the prominent cross-reactivity was observed between the immune sera and R. rickettsii （ ≥1：40） while the little cross-reactivity was observed between the immune sera and typhus group rickettsiae （ ≤1 ： 40）. The results suggest that the recombinant OmpB proteins have a good antigenicity since they have capability to efficiently elicit humoral immunoresponses and to specifically react with rickettsial antibodies.

关 键 词：黑龙江立克次体 外膜蛋白 抗原性 

分 类 号：Q786[生物学—分子生物学]