| 旋毛虫T668基因真核表达载体的构建及其在BHK细胞中的表达 |
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| 引用本文： | 白雪,吴秀萍,王学林,王鑫蕊,于建立,唐斌,刘国兴,冯爽,陈根元,刘明远. 旋毛虫T668基因真核表达载体的构建及其在BHK细胞中的表达[J]. 中国兽医学报, 2010, 30(4) |
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| 作者姓名： | 白雪 吴秀萍 王学林 王鑫蕊 于建立 唐斌 刘国兴 冯爽 陈根元 刘明远 |
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| 作者单位： | 吉林大学,人兽共患病研究所,吉林,长春,130062 |
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| 基金项目： | 国家杰出青年基金，国家科技传染病重大专项，国家自然科学基金 |
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| 摘 要： | 利用RT-PCR技术从中国河南猪旋毛虫(国际标准虫种编号为ISS534)新生幼虫得到T668基因,并克隆入pEGFP-N1真核表达载体中构建重组质粒,该重组质粒在脂质体介导下转染BHK细胞。EGFP标签证明质粒DNA成功转染到细胞中并得以表达,Western blotting鉴定,细胞裂解液样品中有1条约77 000的条带,可被绿色荧光抗体及猪感染旋毛虫阳性血清所识别,与预计大小一致,表明成功构建T668-pEGFP-N1真核表达质粒,并在BHK细胞融合表达,表达产物具有抗原性。
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| 关 键 词： | 旋毛虫 T668基因 真核表达 免疫荧光 |
Construction and expression in BHK cells of eukaryotic expression plasmid with T668 gene of Trichinella spiralis |
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| BAI Xue,WU Xiu-ping,WANG Xue-lin,WANG Xin-rui,YU Jian-li,TANG Bin,LIU Guo-xing,FENG Shuang,CHEN Gen-yuan,LIU Ming-yuan. Construction and expression in BHK cells of eukaryotic expression plasmid with T668 gene of Trichinella spiralis[J]. Chinese Journal of Veterinary Science, 2010, 30(4) |
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| Authors: | BAI Xue WU Xiu-ping WANG Xue-lin WANG Xin-rui YU Jian-li TANG Bin LIU Guo-xing FENG Shuang CHEN Gen-yuan LIU Ming-yuan |
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| Abstract: | The T668 gene was amplified by RT- PCR from Trichinella spiralis NBL originated from pigs in Henan Province ISS534 and cloned into the eukaryotic expression plasmid pEGFP-N1.Then,the pEGFP-N1-T668 plasmid was transfected into BHK cells mediated by GenEscort~(TM) Ⅱ.EGFP showed that the recombinant plasmid was ex-pressed in BHK cells.Western-blotting results illustrated that there was a bands approximately in size in BHK221 cells lysate,it was recognized by the the anti-EGE P antibodies and the pig serum against T.spiralis,indicating that the T668 protein fusion expressed transiently in the cells and had antigenicity. |
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| Keywords: | Trichinella spiralis T668 gene eukaryotic expression Immunofluorescence |
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