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抗汉坦病毒NP scFv基因表达及生物活性分析
引用本文:刘兵,史俊岩,王舰,王美莲,王斯,郑兰艳,牟玲,罗恩杰.抗汉坦病毒NP scFv基因表达及生物活性分析[J].微生物学杂志,2009,29(2):29-32.
作者姓名:刘兵  史俊岩  王舰  王美莲  王斯  郑兰艳  牟玲  罗恩杰
作者单位:1. 中国医科大学,基础医学院,病原生物教研室,辽宁,沈阳,110001
2. 沈阳市传染病院,肾综合征出血热研究所,辽宁,沈阳,110003
摘    要:诱导已构建的重组质粒pGEX-6P—1—scFv原核表达抗汉坦病毒核衣壳蛋白单链抗体,并用酶免疫实验检测单链抗体生物活性。用IPTG诱导重组原核表达质粒pGEX-6P-1-scFv表达抗汉坦病毒NP单链抗体融合蛋白,经亲和层析纯化,并应用SDS—PAGE电泳检测单链抗体融合蛋白,应用酶免疫实验检测抗NP单链抗体生物学活性。SDS—PAGE电泳检测显示,原核重组质粒pGEX-6P-1-scFv已表达分子量约为56ku的单链抗体融合蛋白;酶免疫实验检测显示,单链抗体具有与汉坦病毒NP抗原特异性结合的生物学活性。结果表明,已构建的原核表达重组质粒pGEX-6P-1-scFv,能够成功表达具有与汉坦病毒NP抗原特异性结合生物学活性的单链抗体。

关 键 词:汉坦病毒  scFv  表达

Gene Expression and Bio-Activity Analysis of scFv of Hantavirus NP
LIU Bing,SHI Jun-yan,WANG Jian,WANG Mei-lian,WANG Si,ZHENG Lan-yan,MU Ling,LUO En-jie.Gene Expression and Bio-Activity Analysis of scFv of Hantavirus NP[J].Journal of Microbiology,2009,29(2):29-32.
Authors:LIU Bing  SHI Jun-yan  WANG Jian  WANG Mei-lian  WANG Si  ZHENG Lan-yan  MU Ling  LUO En-jie
Institution:LIU Bing ,SHI Jun-yan, WANG Jian , WANG Mei-lian, WANG Si, ZHENG Lan-yan , MU Ling ,LUO En-jie (1. Res. & Teach. Div. of Pathol. Biol. , Coll. of Basic Med. , China Med. Univ. , Shenyang110001 2. Inst. of Hemorrhagic Fever with Renal Syndrome, Shenyang Infectious Hosp. , Shenyang 110003 )
Abstract:A constructed recombinant plasmid pGEX-6P-1- seFv was induced to express prokaryotie single-chain fragment of variable region (scFv) against nueleocapsid protein (NP) of Hantavirus and to test the bio-aetivity of scFv with immunoassay. IPTG was used to induce the recombinant that expressed prokaryotie plasmid pGEX-6P-1- seFv to express the fusion protein of NP seFv against Hantavirus. After purified by affinity and chromatography, the fusion seFv was tested with SDS-PAGE, bio-aetivity of anti-NP scFv was tested with immunoassay. The results of SDS-PAGE showed that prokaryotie recombinant plasmid pGEX-6P-1- seFv had expressed molecular mass of 56 ku of fusion protein of seFv, and immunoassay test showed that scFv possessed bio-aetivity of ability of specific binding to NP of Han- tavirus. The results showed that the prokaryotic recombinant plasmid pGEX-6P-1- seFv could successfully express the seFv possessing the ability of specific binding to NP of Hantavirus.
Keywords:scFv
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