重组诺如病毒P颗粒的表达纯化及免疫原性

吉林大学学报(理学版)

重组诺如病毒P颗粒的表达纯化及免疫原性

金浩 ¹, 付璐², 史晶莹¹, 吴丛梅¹, 吴慧², 殷玉和¹

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

收稿日期:2016-10-08 出版日期:2017-07-26 发布日期:2017-07-13
通讯作者: 殷玉和 E-mail:yyhlxt72@163.com

Expression, Purification and Immunogenicity of Recombinant Norovirus P Particles

JIN Hao¹, FU Lu², SHI Jingying ¹, WU Congmei ¹, WU Hui², YIN Yuhe ¹

1. College of Chemistry and Life Science, Changchun University of Technology, Changchun 130012, China;2. College of Life Sciences, Jilin University, Changchun 130012, China

Received:2016-10-08 Online:2017-07-26 Published:2017-07-13
Contact: YIN Yuhe E-mail:yyhlxt72@163.com

摘要/Abstract

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Native-PAGE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20 nm, 并对β淀粉样蛋白有免疫反应.

关键词: 诺如病毒重组P颗粒, 蛋白免疫原性, 可溶性表达, 蛋白疫苗

Abstract: 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 are 24mer, 12mer and 2mer, and the particle size of the protein is about 20 nm, and has an immune response to the βamyloid protein.

Key words: Norovirus recombinant P particle, soluble expression; protein immunogenicity, protein vaccine

中图分类号:

Q819

金浩, 付璐, 史晶莹, 吴丛梅, 吴慧, 殷玉和. 重组诺如病毒P颗粒的表达纯化及免疫原性[J]. 吉林大学学报(理学版), 2017, 55(04): 1025-1030.

JIN Hao, FU Lu, SHI Jingying, WU Congmei, WU Hui, YIN Yuhe. Expression, Purification and Immunogenicity of Recombinant Norovirus P Particles[J]. Journal of Jilin University Science Edition, 2017, 55(04): 1025-1030.