吕氏泰勒虫TlSP基因在真核细胞中的表达与定位  

Expression and localization of TlSP gene of Theileria luwenshuni in eukaryotic cells

作　　者：何海宁[1] 刘志杰[2] 杨吉飞[2] 陈泽[2] 关贵全[2] 王雅[1] 刘爱红[2] 任巧云[2] 罗建勋[2] 殷宏[2] 赵萍[1] 李有全[2] 

机构地区：[1]兰州理工大学生命科学与工程学院,甘肃兰州730050 [2]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室甘肃省动物寄生虫病重点实验室,甘肃兰州730046

出　　处：《中国兽医科学》2013年第9期919-923,共5页Chinese Veterinary Science

基　　金：国家自然科学基金资助项目(31272556;30972182;31072130;31001061);PIROVAC项目(KBBE-3-245145);国家重点基础研究发展计划(973)项目(2010CB530206);国家引进国际先进农业科学技术计划(948)项目(2010-S04);甘肃省重点项目(1002NKDA035);家畜疫病病原生物学国家重点实验室项目(SKLVEB2008ZZKT019);欧盟EPIZONE(FOOD-CT-2006-016236);ASFRISK(211691);ARBOZOONET(211757)

摘　　要：为研究吕氏泰勒虫TlSP基因在真核细胞中的表达与亚细胞定位,构建了pEGFP-N1-TlSP重组表达质粒,经脂质体转染Vero细胞后,用Western-blot验证了该蛋白在Vero细胞的表达;用荧光显微镜和激光共聚焦显微成像技术确定了TlSP重组蛋白在真核细胞中的分布及亚细胞定位。结果表明重组质粒pEGFP-N1-TlSP在Vero细胞中获得了表达,绿色荧光主要聚集于细胞浆,而在空载体pEGFP-N1转染组,绿色荧光弥散性分布于细胞核。吕氏泰勒虫TlSP融合基因在外源真核细胞Vero中成功进行了表达,表达蛋白主要定位于Vero细胞的细胞浆。The aim of this study was to study the expression and subcellular localization of T1SP gene of Theileria luwenshuni in eukaryotic cells. The pEGFP-N1-T1SP recombinant plasmid was constructed and transfected into Vero cells with lipofectamine 2000. Western-blot was used to verify the expression of rT1SP recombinant protein. The distribution and subcellular localization of rT1SP recombinant protein in eukaryotic cells was determined using fluorescent microscope and laser confocal microscopy imaging tech- nique. The results showed that the recombinant plasmid was successfully expressed in Vero cells and the green fluorescence was distributed in the cytoplasm;however, the green fluorescence distributed in nucleus in empty vector. Therefore, T. luwenshuni T1SP gene was expressed successfully in heterologous eukaryotic Vero cells and the expression of protein mainly located in Veto cell cytoplasm.

关 键 词：吕氏泰勒虫 TlSP基因 绿色荧光蛋白 激光共聚焦显微成像技术 

分 类 号：S852.723[农业科学—基础兽医学]