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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 study the immunogenicity and tissue localization of the recombinant protein of Annexin B3 (B3) derived from Taenia asiatica. Methods Annexin B3 gene was constructed and expressed in prokaryotic expression plasmid pET-28a (+) in E. coli BL21 / DE3. The purified recombinant protein was analyzed by Western blotting and indirect Fluorescent immunohistochemical localization and observation under a fluorescence microscope. Results The induced recombinant protein was purified and identified by the serum of patients infected with Taenia asiatis, Taenia solium and Taenia saginata. Immunohistochemistry showed that the tissue of this gene is localized on the epidermis of tapeworm Asia. Conclusion The recombinant protein has good immunoreactivity and its localization in the epidermis suggests that this gene is a key molecule to maintain the balance of energy metabolism in the anaerobic environment of the host digestive tract and to mediate the potential interaction between parasites and host Vaccine candidate antigen molecule.