日本血吸虫表面膜抗原23kDa的亚克隆鉴定及表达

THE SUBCLONING, VERIFICATION AND EXPRESSION OF SCHISTOSOMA JAPONICUM 23KDA GENE

在已克隆的质粒pBluescript-Sj23基础上，设计二条引物。小量提取质粒pBluescript－Sj23进行PCR扩增，经扩增的Sj23亚克隆于真核表达载体pCD中，重组质粒转化E．coliTG1。大量提取质粒pCD－Sj23，以剂量100μg／只给昆明小白鼠定位肌肉注射质粒pCD-Sj23，共注射2次，间隔时间为9天，每次注射前一天用0．5％盐酸布比卡因50μl／只预处理，第14天拉颈处死小鼠，定位处肌肉作冰冻切片，荧光标记抗体检测pCD-Sj23，见肌细胞胞浆及胞膜上有其抗原表达。

According to the published gene sequence of the Sj23, a pair of primers were designed- With the twoprimers,S j23 isolated from E- coli TG, was amplified and subcloned into eukaryocyte expression vector. Recombinant plasmidwere injected into muscle of mice,each of them was injected twice at the same point at a 9 days interval. The mice were sacri-ficed on the fourteenth day. Using the technology of histochemistry. A Ag-Ab reaction were observed in the frozen musculartissue slice,the resu1t indicated that the antigen cou1d be expressed on the membrane of muscular cells.