旋毛虫Ts43基因真核表达载体的构建及其在Vero E6细胞中的表达  

Construction of eukaryotic expression vector for Ts43 gene from Trichinella spiralis and expression of the gene in Vero E6 cells

作　　者：王君[1] 路义鑫[1] 姜曰晓[1] 朱江威[1] 朱艳梅[1] 袁金钱[1] 宋铭忻[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国兽医科学》2007年第6期500-504,共5页Chinese Veterinary Science

基　　金：霍英东青年教师基金项目(91034);中国科技部自然科技资源基础平台项目(2006BAD06A09);中国博士后科学基金项目(2004035160)

摘　　要：利用RT-PCR技术从黑龙江省猪源旋毛虫获得了Ts43基因,并克隆入pcDNA3.1-CT-GFP真核表达载体中构建重组质粒,用该重组质粒在脂质体介导下转染Vero E6细胞,GFP标签证明质粒DNA成功转染到细胞中并得以表达,通过Western-blot分析,细胞裂解液样品中有1条约66 ku的条带,可被小鼠旋毛虫阳性血清所识别,与预计大小一致,说明,真核表达载体pcDNA3.1-CT-GFP中的Ts43基因在VeroE6细胞中获得了表达,表达产物具有抗原性。The Ts43 gene of Trichinella spiralis originated from pigs in Heilongjiang Province was amplified by RT-PCR and subcloned into pcDNA3. 1-CT-GFP vector to construct recombinant plasmid pcDNA3. 1-CT-GFP-Ts43. Sequence analysis revealed that the Ts43 gene was correctly inserted into the pcDNA3.1-CT-GFP. Then, the pcDNA3.1-CT-GFP-Ts43 plasmid was transfected into Vero E6 cells mediated by Lipofectamine 2000. GFP showed that the recombinant plasmid was expressed in Vero E6 cells. Western-blot analysis showed that a band approximately 66 ku in size in Vero E6 cells lysate was recognized by the mouse serum against T. spiralis, indicating that the Ts43 protein expressed transiently in the cells had antigenicity.

关 键 词：旋毛虫 Ts43基因 克隆 真核表达 

分 类 号：S852.731[农业科学—基础兽医学]