重组诺如病毒P颗粒的表达纯化及免疫原性  

Expression,Purification and Immunogenicity of Recombinant Norovirus P Particles

作　　者：金浩[1] 付璐[2] 史晶莹[1] 吴丛梅[1] 吴慧[2] 殷玉和[1] JIN Hao FU Lu SHI Jingying WU Congmei WU Hui YIN Yuhe(College of Chemistry and Life Science, Changchun University of Technology, Changchun 130012, China College of Life Sciences, Jilin University, Changchun 130012, China)

机构地区：[1]长春工业大学化学与生命科学学院,长春130012 [2]吉林大学生命科学学院,长春130012

出　　处：《吉林大学学报（理学版）》2017年第4期1025-1030,共6页Journal of Jilin University:Science Edition

基　　金：吉林省科技发展计划项目(批准号:20150101226JC)

摘　　要：通过构建重组质粒并表达纯化目的蛋白,用SDS-PAGE、Western blot和NativePAGE检测蛋白,用Superdex^(TM)200凝胶色谱层析分析目的蛋白的多聚体,用动态光散射以及透射电镜对目的蛋白颗粒进行分析,并通过小鼠免疫,用四免血清进行酶联免疫吸附实验检测蛋白的免疫反应.结果表明:成功构建了质粒pET26b-PP-3copy-Aβ1-6-loop123,并成功表达目的蛋白;目的蛋白存在3种寡聚体形式,分别为24聚体、12聚体和二聚体;蛋白颗粒约为20nm,并对β淀粉样蛋白有免疫反应.We constructed a recombinant plasmid and expressed the purified target protein,and used SDS-PAGE,Western blot and Native-PAGE to test protein.The multimers of the target protein were analyzed by Superdex^（TM） 200 gel chromatography,and the particle size of the target protein was analyzed by dynamic light scattering and transmission electron microscopy.The immune response of the protein was detected by enzyme-linked immunosorbent assay with four-immunized serum.The results show that plasmid pET26b-PP-3copy-Aβ1-6-loop123 was successfully constructed,and the target protein was successfully expressed.There are three oligomers in the target protein,which are24-mer,12-mer and 2-mer,and the particle size of the protein is about 20 nm,and has an immune response to theβ-amyloid protein.

关 键 词：诺如病毒重组P颗粒 可溶性表达 蛋白免疫原性 蛋白疫苗 

分 类 号：Q819[生物学—生物工程]