猪囊尾蚴抗原基因TS11的真核表达研究  

The eukaryotic expression of TS11 antigenetic gene of Cysticercus cellulosae

作　　者：沈翼[1] 高荣[1] 沈斌[2] 武梅[1] 孟民杰[1] 唐漫书[1] 李江凌[1] 郑勇[1] 刘崑[1] 魏竹波[1] 王克非[1] 刘世贵[1] 

机构地区：[1]四川大学草原生物防治工程国家专业实验室,四川成都610064 [2]四川华神集团企业技术中心博士后流动站,四川成都610015

出　　处：《中国兽医杂志》2003年第4期6-9,共4页Chinese Journal of Veterinary Medicine

基　　金：高等学校博士学科点专项科研基金资助项目 (9410 0 6)

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。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.

关 键 词：猪囊尾蚴 抗原基因 TS11基因 真核表达 蛋白表达产物 人畜共患病 

分 类 号：S855.99[农业科学—临床兽医学]