抗汉坦病毒NP scFv基因表达及生物活性分析  

Gene Expression and Bio-Activity Analysis of scFv of Hantavirus NP

作　　者：刘兵[1] 史俊岩[1] 王舰[1] 王美莲[1] 王斯[1] 郑兰艳[1] 牟玲[2] 罗恩杰[1] 

机构地区：[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

出　　处：《微生物学杂志》2009年第2期29-32,共4页Journal of Microbiology

基　　金：辽宁省自然科学基金资助(20062101)

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。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.

关 键 词：汉坦病毒 SCFV 表达 

分 类 号：R373.3[医药卫生—病原生物学]