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  

High Level Expression of Fusion Protein Gene of Newcastle Disease Virus in E.coli and Analysis of its Immunogenicity

作　　者：游猛[1] 潘志明[1] 耿士忠[1] 文科[1] 陈俊华[1] 张磊[1] 方强[1] 蔡雯婷[1] 焦新安[1] 

机构地区：[1]扬州大学江苏省人兽共患病学重点实验室,江苏扬州225009

出　　处：《中国家禽》2009年第20期21-24,共4页China Poultry

基　　金：国家自然科学基金资助项目(30871860);高等学校科技创新工程重大项目培育资金项目(706032);江苏省自然科学基金项目(BK2007511);江苏省高校自然科学基础研究项目(07KJB230136)

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。A part of the fusion protein （F） gene which is 594 bp of Newcastle disease virus was amplified by polymerase chain reaction （PCR） from /he 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.

关 键 词：新城疫病毒 融合蛋白 原核表达 免疫原性 

分 类 号：S852.659.4[农业科学—基础兽医学] Q78[农业科学—兽医学]