【摘要】 将猪囊尾蚴抗原基因 TS11亚克隆至 VR10 2 0真核表达载体中 ,用菌落原位杂交法筛选重组质粒 ,将其转染 COS7细胞 ,以 Northern Blotting检测插入片段的转录 ;用 EL ISA检查其翻译表达产物。结果表明 :VTS11在真核细胞中能够转录TS11基因的 m RNA;其蛋白表达产物能为兔抗猪囊虫高免血清所识别 ,在转染真核细胞 2 4 h后 ,即可检测到正确表达的猪囊尾蚴抗原蛋白。这些为深入研制猪囊虫的 DNA疫苗奠定了可靠的基础。更多还原

【Abstract】 VTS11, a eukaryotic expression recombinant plasmid of an antigenetic gene of Cysticercus cellulosae was constructed, VTS11 was transformed into E.coli DH5α competent cells. The recombinant plasmid were extracted and digested with Sacl and Pstl to identified the correct orientation. VTS11 plasmid DNA was entrapped with Lipofectamine TM Reagen and expressed in COS7 cells in vitro. The expressed products of VTS11 in COS7 cells were analyzed by ELISA with rabbit antisera against cysticercus in 24 and 72 hours. mRNA of the transfected COS7 cells was extracted and detected by Northern blotting. Results showed that VTS11 could express full-length antigen in vitro in COS7 cells. A transcript on the Northern blot that comes only from the recombinant plasmid containing TS11 gene in COS7 cells. So the plasmid VTS11 encoding immunogenic protein gene of Cysticercus cellulosae could be employed as a candidate for DNA vaccination against Taenia solium crysticerosis.更多还原