中国兽医科学2023，Vol.53Issue(12)：1507-1515,9.DOI:10.16656/j.issn.1673-4696.2023.0210
A型口蹄疫病毒AF72株病毒样颗粒制备条件优化及鉴定
Optimization of preparation conditions and identification of virus-like particles of AF72 strain of foot-and-mouth disease virus type A
摘要
Abstract
To prepare virus-like particles of AF72 strain of FMDV type A,this study improved the soluble expression of the target protein by optimizing IPTG concentration,D600 value of bacterial solu-tion at induction,induction temperature,and induction time.The particles were assembled in a specific enzyme-cut assembly solution and the composition of the assembly solution was adjusted to improve its assembly rate in vitro.Subsequently,SDS-PAGE,sucrose density gradient centrifugation,dynamic light scattering,DAS-ELISA,and transmission electron microscopy were used to determine target protein.The results showed that the three bands of VP0,VP1 and VP3 were identified by SDS-PAGE and were consistent with the size of the target band.The particle size of the nanoparticle was about 30 nm and was consis-tent with the size of the virus particles of foot-and-mouth disease.VP0,VP1 and VP3 was identified by DAS-ELISA and successfully assembled into virus-like particles in vi tro,and some virus-like particles were also photographed by electron microscope.The above results indicated that virus-like particle of AF72 strain of FMDV type A were successfully prepared,which laid a foundation for establishing a liquid-phase blocking ELISA for detecting FMDV neutralizing antibody.关键词
A型口蹄疫/AF72株/VP0、VP1、VP3融合表达/病毒样颗粒组装鉴定Key words
FMDV type A/AF72 strain/VP0,VP1 and VP3 fusion expression/identification of virus-like particle assembly分类
农业科技引用本文复制引用
张雅雅,敬晓棋,李昊洲,王运航,王田田,李河林,黄增文,孙世琪,郭慧琛,毕台飞..A型口蹄疫病毒AF72株病毒样颗粒制备条件优化及鉴定[J].中国兽医科学,2023,53(12):1507-1515,9.基金项目
榆林市产学研科技项目(CXY-2021-87) （CXY-2021-87）
陕西省科技厅一般项目(S2023-YF-YBNY-0080) （S2023-YF-YBNY-0080）
陕西省重点研发计划项目(2023-YBNY-143) （2023-YBNY-143）
