A型塞内卡病毒VP2蛋白原核表达及免疫原性分析  

Prokaryotic Expression and Immunogenicity of Senecavirus A VP2 Protein

作　　者：崔川 刘佳欢 韩伟建 张俊娟[1] 张义明[1] 李丽敏[1] CUI Chuan;LIU Jiahuan;HAN Weijian;ZHANG Junjuan;ZHANG Yiming;LI Limin(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071000,China)

机构地区：[1]河北农业大学动物医学院,保定071000

出　　处：《中国动物传染病学报》2022年第6期91-98,共8页Chinese Journal of Animal Infectious Diseases

基　　金：河北省第三批青年拔尖人才计划;河北农业大学科研发展计划(JY2019045)。

摘　　要：为研究A型塞内卡病毒(SVA)VP2蛋白的生物学特性,本试验对GenBank公布的SVA VP2基因序列分析筛选,合成完整VP2基因序列。使用Expasy软件对其结构进行分析,通过原核表达VP2蛋白并利用正交设计优化其可溶性表达条件,纯化VP2蛋白免疫獭兔、小鼠,检测血清中特异性抗体和细胞因子的变化。结果显示:SVA VP2蛋白的一、二级结构得以展示;成功表达了可溶性的VP2蛋白,可溶性蛋白表达最佳条件为培养转速120 rpm、诱导时间8 h、诱导温度30℃、诱导剂IPTG浓度0.5 mmol/L;制备了高效价的兔源多克隆抗体;在免疫VP2蛋白的小鼠血清中检测到高水平的VP2特异性抗体且IFN-γ和IL-2水平显著升高,表明VP2蛋白具有良好的免疫原性。本研究为进一步塞内卡病毒VP2蛋白功能及应用研究提供了技术支持。To study the biological characteristics of SVA VP2 protein, the SVA VP2 gene sequences published in GenBank were analyzed and the complete VP2 gene was synthesized in our study. The structure of VP2 protein was analyzed by expasy software and its soluble prokaryotic expression conditions were optimized by orthogonal design. The expressed VP2 protein was purifi ed and used to immunize rex rabbits and mice, and the levels of specifi c antibodies and cytokines in serum were detected. The results showed that the soluble SVA VP2 protein was successfully expressed so that the primary and secondary structures of VP2 protein were able to be demonstrated. The soluble protein expression conditions were optimized as culture speed at 120 rpm, induction time for 8 h and temperature at 30℃, and inducer IPTG concentration of 0.5 mmol/L. The rabbit polyclonal antibodies were prepared with high titer. High level of VP2 specifi c antibodies was detected in the mouse serum and the levels of IFN-γ and IL-2 were signifi cantly increased, which indicated that VP2protein had good immunogenicity. This study provided technical support for further study of the function and application of SVA VP2protein.

关 键 词：A型塞内卡病毒 VP2基因 免疫原性 细胞因子 正交设计 

分 类 号：S852.4[农业科学—基础兽医学]