含FMDV中和表位的重组病毒颗粒的构建及鉴定  

Construction and Identification of Recombinant Virus-like Particles Fused with Neutralizing Epitopes of FMDV

作　　者：苗巍男 李瑞[1] 钟江[1] 郑兆鑫[1] 刘明秋[1] 

机构地区：[1]复旦大学生命科学学院,上海200438

出　　处：《复旦学报（自然科学版）》2016年第3期381-387,共7页Journal of Fudan University：Natural Science

基　　金：教育部留学回国人员科研启动基金;上海市科学技术委员会项目(13DZ2252000)

摘　　要：针对FMDV的疫苗研究一直是防控口蹄疫的重点,而病毒样颗粒（VLPs）形式的疫苗因其安全性和近似传统灭活疫苗的良好免疫效果,近年来逐渐成为疫苗研究的一个新方向.本实验利用猪圆环病毒PCV2的衣壳蛋白（CAP）作为载体,分别插入FMDV MYA98毒株的抗原表位组合——VP1蛋白上B细胞表位141～160aa（B）与串联的B细胞和T细胞表位141～160aa-21～40aa-141～160aa（BTB）,构建对应重组杆状病毒,分别命名为reBAC-CAP-B,reBAC-CAP-BTB.将上述病毒颗粒转入Sf9细胞表达,收获重组杆状病毒并再次感染Sf9细胞扩大病毒滴度,获得目的蛋白CAP-B,CAP-BTB,经过SDS-PAGE,Western blot鉴定正确,透射电镜观察到20～30nm大小的目的颗粒.The study on vaccine of FMDV is always a hot spot in world. Recent years, VLPs became a new direction of vaccine study, for its safety and good ability of protection against virus compared to traditional vaccines. Two kinds of neutralizing epitopes of type O FMDV MYA98, single B antigen epitope on VP1 141- 160 aa（B） and the combination of B antigen epitope and T antigen epitope 141-160 aa-21-40 aa-141-160 aa （BTB） epitopes, were inserted into CAP vector protein from PCV2 to construct the recombined Bacmids reBAC- CAP-B, reBAC-CAP-BTB. These recombined Bacmids were transfected into Sf9 cells to express fusion proteins CAP-B,CAP-BTB respectively. After amplifying the viral stock, the high-titer stock was infected to cells for protein expression. The results of SDS-PAGE, Western blot and TEM showed that the proteins with the right molecule size had the specific immunogenicity and were assembled into VLPs successfully.

关 键 词：口蹄疫病毒 猪圆环病毒 病毒样颗粒 抗原表位 杆状病毒-昆虫细胞表达系统 

分 类 号：Q78[生物学—分子生物学]