| 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 |
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| 引用本文： | 王君,路义鑫,姜曰晓,朱江威,朱艳梅,袁金钱,宋铭忻.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[J].中国兽医科学,2007,37(6):500-504. |
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| 作者姓名： | 王君 路义鑫 姜曰晓 朱江威 朱艳梅 袁金钱 宋铭忻 |
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| 作者单位： | 东北农业大学,动物医学学院,黑龙江,哈尔滨,150030 |
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| 基金项目： | 霍英东教育基金;科技部科技基础条件平台建设计划;中国博士后科学基金 |
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| 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。
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| 关 键 词： | 旋毛虫 Ts43基因 克隆 真核表达 |
| 文章编号： | 1673-4696(2007)06-0500-05 |
| 修稿时间： | 2007年1月16日 |
Construction of eukaryotic expression vector for Ts43 gene from Trichinella spiralis and expression of the gene in Vero E6 cells |
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| WANG Jun,LU Yi-xin,JIANG Yue-xiao,ZHU Jiang-wei,ZHU Yan-mei,YUAN Jin-qian,SONG Ming-xin.Construction of eukaryotic expression vector for Ts43 gene from Trichinella spiralis and expression of the gene in Vero E6 cells[J].Veterinary Science in China,2007,37(6):500-504. |
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| Authors: | WANG Jun LU Yi-xin JIANG Yue-xiao ZHU Jiang-wei ZHU Yan-mei YUAN Jin-qian SONG Ming-xin |
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| Abstract: | 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. |
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| Keywords: | Trichinella spiralis Ts43 gene cloning eukaryotic expression |
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