猪囊虫抗原基因TS21的真核表达

The eukaryotic expression of TS21 antigenetic gene of Taenia solium

构建了猪囊虫抗原基因TS2 1的VR10 2 0真核表达载体 ,以菌落原位杂交法筛选获得正确插入的重组质粒 ,用RNA印迹 (Northernblotting)和ELISA检测插入片段的转录及其翻译表达产物。结果 ,VTS2 1在真核细胞中能够转录TS2 1基因的mRNA ;兔抗猪囊虫超免疫血清可识别其蛋白表达产物 ,表明VTS2

The experiment was conducted to construct VTS21, a eukaryotic expression recombinant plasmidof an antigenetic gene of Taenia solium. VTS21 was transformed into E.coli DH5α competent cells. The recombinant plasmid were extracted and digested incompletely with XmaⅢ to identify the correct orientation. VTS21 plasmid DNA was entrapped with Lipofectamine TM Reagentand expressed in Cos7 cells. The expressedproducts of VTS21 in Cos7 cells were analyzed by ELISA with rabbit antisera against cysticercus, mRNA of the transfected Cos7 cells was extracted and detected by Northern blotting. Results indicated that VTS21 can express full length antigen in vitro in Cos7 cells, and that a transcript observed on the Northern blottingcomes only from the recombinant plasmid containing TS21 gene in Cos7 cells. So the VTS21 plasmid encoding the immunogenic protein of Taenia solium can be employed as a candidate for DNA vaccine against Taenia solium cysticerosis.