含牛轮状病毒VP6的嵌合型HBcAg颗粒抗原的制备及其免疫原性分析  

Prokaryotic expression and immunogenicity of the chimer HBcAg containing bovine rotavirus VP6

作　　者：邢亚茹 张增贤 顾文源[1,3] 郭禹 彭志豪 左玉柱[1,4] 范京惠 XING Yaru;ZHANG Zengxian;GU Wenyuan;GUO Yu;PENG Zhihao;ZUO Yuzhu;FAN Jinghui(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071001,China;Ningjin Agricultural and Rural Bureau,Ningjin 055550,China;Hebei Disease Control and Prevention Center,Shijiazhuang 050035,China;Hebei Veterinary Biotechnology Innovation Center,Baoding 071001,China)

机构地区：[1]河北农业大学动物医学院,河北保定071001 [2]宁晋县农业农村局,河北宁晋055550 [3]河北省动物疫病预防控制中心,河北石家庄050035 [4]河北省兽医生物技术创新中心,河北保定071001

出　　处：《河北农业大学学报》2021年第3期79-84,共6页Journal of Hebei Agricultural University

基　　金：河北农业大学精准畜牧学科群建设项目(1090064);河北省农业产业技术体系奶牛创新团队项目(HBCT2018120406);河北省农业产业技术体系肉牛创新团队项目(HBCT2018130405).

摘　　要：本研究将编码牛轮状病毒VP6蛋白的部分基因插入到HBcAg的免疫显性区的基因中,构建了pET32a-HBcAg-VP6原核表达载体,并将其转化至大肠杆菌BL21中进行表达。重组蛋白经纯化和Western blot鉴定后,通过透射电镜观察重组蛋白HBc-VP6是否形成病毒样颗粒。以重组蛋白HBc-VP6皮下注射BALB/c小鼠,对免疫原性进行初步评价。结果显示:重组蛋白HBc-VP6在BL21中主要以沉淀的形式表达,经纯化、复性后的重组蛋白HBc-VP6能够自我折叠装配成病毒样颗粒结构,能够与BRV阳性血清发生特异性反应。重组蛋白HBc-VP6能够刺激小鼠产生抗BRV特异性抗体,同时可以产生相关的Th1型和Th2型细胞因子,试验结果证实,本研究成功制备了含VP6和HBcAg的重组蛋白HBc-VP6,并在小鼠模型中初步评价了其免疫原性,为今后牛轮状病毒疫苗的研制提供了新的思路。In this study,a part of the gene encoding the bovine rotavirus VP6 protein was inserted into the immunodominant region of HBcAg to construct the prokaryotic expression vector pET32a-HBcAg-VP6,which was transformed into E.coli BL21.After purification and identification by Western-blot,the transmission electron microscope was used to observe whether the recombinant protein HBc-VP6 forms virus-like particles.The expressed recombinant proteins HBc-VP6 were injected into BALB/c mice as vaccine antigen through subcutaneous injection and its immunogenicity were preliminary evaluated.The results showed that the recombinant protein HBc-VP6 was mainly expressed in the form of precipitation in BL21.The purified and renatured recombinant protein HBc-VP6 can self-fold and assemble into a virus-like particle structure,which can react specifically with BRV positive serum.Besides,the HBc-VP6 could induce anti-PEDV specific antibodies and the related Th1 and Th2 cytokines in mice.The results showed that this study successfully prepared the recombinant protein HBc-VP6 containing VP6 and HBcAg,and initially evaluated its immunogenicity in mice.The results of this study provides new idea for the development of bovine rotavirus vaccines in the future.

关 键 词：牛轮状病毒 乙肝病毒核心抗原 病毒样颗粒 VP6基因 

分 类 号：S852.653[农业科学—基础兽医学]