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目的 获得旋毛虫成虫抗原基因的原核表达产物并对其进行纯化及免疫特性分析。 方法 旋毛虫成虫Ts87基因片段亚克隆入 PET- 2 8a(+)表达载体 ,异丙基硫代 -β- D-半乳糖苷 (IPTG)诱导表达。亲和层析和电洗脱法纯化表达产物 ,SDS- PAGE、 EL ISA和 Western blotting对表达产物进行分析鉴定 ,并将纯化的表达产物人工免疫兔。结果 SDS- PAGE结果显示 ,表达产物为分子量约为 4 0 k Da的重组蛋白。纯化后免疫兔获得高滴度的抗血清。该基因重组蛋白可被感染旋毛虫的猪、兔血清及人工免疫兔血清所识别 ,与感染囊尾蚴、棘球蚴的患者血清或感染日本血吸虫的兔血清不发生交叉免疫反应。 结论 获得一个新的旋毛虫成虫 Ts87基因重组蛋白 ,该蛋白具有特异的抗原性。
Objective To obtain the prokaryotic expression product of the Trichinella spiralis adult antigen gene and to analyze the purification and immunological characteristics. Methods The Ts87 gene of Trichinella spiralis was subcloned into the expression vector PET-2 8a (+) and induced by isopropyl thio-β-D-galactoside (IPTG). The expressed products were purified by affinity chromatography and electroelution. The expressed products were identified by SDS-PAGE, ELISA and Western blotting. The expressed products were artificially immunized with rabbit. Results The results of SDS-PAGE showed that the expressed product was a recombinant protein with a molecular weight of about 40 kDa. After purification, rabbits were immunized to obtain high titer antisera. The recombinant protein of this gene can be recognized by the trichinella swine, rabbit serum and artificial immune rabbit serum, and does not cross-react with serum of patients infected with cysticercosis, hydatid cyst, or rabbit serum infected with Schistosoma japonicum. Conclusion A new Trichinella spiralis Ts87 gene recombinant protein was obtained, which has specific antigenicity.