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当前位置: 首页 > 文章 > 新城疫病毒F基因在大肠杆菌中的高效表达及其免疫原性分析 中国家禽 2009,31 (20) 21-24
Position: Home > Articles > High Level Expression of Fusion Protein Gene of Newcastle Disease Virus in E.coli and Analysis of its Immunogenicity China Poultry 2009,31 (20) 21-24

新城疫病毒F基因在大肠杆菌中的高效表达及其免疫原性分析

作  者：
游猛;潘志明;耿士忠;文科;陈俊华;张磊;方强;蔡雯婷;焦新安
单  位：
扬州大学江苏省人兽共患病学重点实验室
关键词：
新城疫病毒;融合蛋白;原核表达;免疫原性
摘  要：
应用PCR技术,以含有新城疫病毒F48E8株全长融合蛋白(F)基因的真核表达质粒pVAX1-F为模板,扩增出594bp大小的F基因的部分片段。经克隆筛选和测序后,构建成重组原核表达质粒pGEX-6P-1-F。重组质粒转化表达菌BL21,获得重组菌BL21(pGEX-6P-1-F)。经IPTG诱导和SDS-PAGE分析表明,F基因在原核表达体系中获得高效表达,表达量占菌体总蛋白的23%。Western blotting结果显示,在相对分子质量46ku位置有特异性条带。动物实验结果表明,F蛋白免疫鸡产生的针对新城疫病毒F蛋白的血清抗体效价显著高于空白对照组(P<0.05),说明原核表达系统表达的F蛋白具有良好的免疫原性。
译  名：
High Level Expression of Fusion Protein Gene of Newcastle Disease Virus in E.coli and Analysis of its Immunogenicity
作  者：
YOU Meng,PAN Zhiming,GENG Shizhong,WEN Ke,CHEN Junhua, ZHANG Lei,FANG Qiang,CAI Wenting,JIAO Xin′an (Jiangsu Key Laboratory of Zoonosis,Yangzhou University,Yangzhou,Jiangsu 225009)
关键词：
Newcastle disease virus;fusion protein;prokaryotic expression;immunogenicity
摘  要：
A part of the fusion protein (F) gene which is 594 bp of Newcastle disease virus was amplified by polymerase chain reaction (PCR) from the recombinant eukaryotic expression plasmid pVAX1 -F,and subcloned into prokaryotic expression vector pGEX-6P-1. The F gene was identified by sequencing. Then the recombinant plasmid was transformed into E.coli BL21, and the recombinant was designated as BL21 (pGEX-6P-1-F). The result of SDS-PAGE verified that there was a desired recombinant protein with molecular weight of 46 000 and accounted for 23% of the total bacterial protein. Western blotting analysis further indicated this expressed product had the immunogenicity of F protein. The F protein could significantly elicit specific serum immune response in immunized SPF chickens,compared with blank control (P<0.05). These results demonstrated that the F protein expressed by E.coli has good immunogenicity.

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