亚洲带绦虫膜联蛋白B3基因免疫学特性分析及组织定位  

The expression and purification of the Annexins B3 gene in Taenia saginata asiatica and the immunological analysis of the recombinant protein

作　　者：戴佳琳[1] 蓝磊[2] 黄江[1] 

机构地区：[1]贵阳医学院法医学系,贵州贵阳550004 [2]贵阳市妇幼保健院,贵州贵阳550004

出　　处：《热带医学杂志》2014年第8期1009-1011,F0003,共4页Journal of Tropical Medicine

基　　金：国家自然科学基金(81260256)

摘　　要：目的对重组表达的亚洲带绦虫膜联蛋白B3(Annexins B3)基因得到的蛋白进行免疫学初步研究及组织定位。方法将构建在原核表达质粒pET-28a(+)中的Annexins B3基因在大肠埃希菌BL21/DE3中进行诱导表达,将纯化后的重组蛋白用蛋白印迹(Western blotting)进行免疫学分析,间接荧光免疫组织化学定位,荧光显微镜下曝光观察结果。结果诱导表达并纯化后的重组蛋白可被亚洲带绦虫、猪带绦虫和牛带绦虫感染的病人血清识别。免疫组化显示,该基因的组织定位在亚洲带绦虫成虫的表膜。结论该重组蛋白具有较好的免疫反应性,组织定位于表膜,说明该基因是维持成虫在宿主消化道厌氧环境中能量代谢平衡的关键分子和介导寄生虫与宿主相互作用的潜在的疫苗候选抗原分子。Objective To analysis the immunogenicity and tissue localization of the Annexins B3 recombinant protein of Taenia saginata asiatica. Methods The recombinant protein was detected by SDS-PAGE after being induced with IPTG in E.coli BL21 / DE3 and purified with NI-IDA affinity chromatography successfully. In addition, the immunoreactivity of the purified recombinant proteins was analyzed by Western Blotting. The localization of Ts Annexins B3 protein in adult worms was demonstrated by immunofluorescent technique. Results Western blot analysis showed that the serum from patients infected with T.saginata, T. asiatica could react with the purified recombinant protein. The immunofluorescence assay revealed that the Annexins B3 protein was located at the tegument of Taenia saginata asitica adult. Conclusion The purified recombinant protein of the Annexins B3 was immunogenic. The protein is located at the outer membrane of the tegument of the adult worm.It could be the crucial molecule involving in the maintenance of the energy metabolism and balance of the adult dwelling in a hypoxia environment in host intestine duct, and they could also be the potential vaccine candidate.

关 键 词：亚洲带绦虫 膜联蛋白B3 免疫反应性 组织定位 

分 类 号：R383.32[医药卫生—医学寄生虫学]