携带猪流行性腹泻病毒S1抗原表位的乙肝核心抗原病毒样颗粒的构建与鉴定  

Construction and Identification of Hepatitis B Core Antigen Virus-like Particles Carrying PEDV S1 Epitope

作　　者：刘如月 范京惠[1] 刘涛[2] 苑军辉 李清艳[1] 翟向和[1] 左玉柱[1] LIU Ruyue;FAN Jinghui;LIU Tao;YUAN Junhui;LI Qingyan;ZHAI Xianghe;ZUO Yuzhu(College of Veterinary Medicine/Agriculture Science and Education Center,Hebei Agricultural University,Baoding 071001,China;Ringpu(Baoding)Biological Pharmaceutical Co.Ltd.,Baoding 071001,China;Xingtang Animal Health Supervision Institution,Xingtang 050600,China)

机构地区：[1]河北农业大学动物医学院/科教兴农中心,保定071001 [2]瑞普(保定)生物药业有限公司,保定071001 [3]行唐县动物卫生监督所,行唐050600

出　　处：《畜牧兽医学报》2021年第6期1760-1764,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：河北省重点研发计划项目(19226622D):河北省农业产业技术体系生猪创新团队(HBCT2018110207)。

摘　　要：旨在构建以乙肝核心抗原(HBcAg)为载体呈现猪流行性腹泻病毒(PEDV)S1抗原表位的病毒样颗粒。将PEDV S1蛋白中含B细胞表位的270 bp片段插入到HBcAg主要免疫显性区域(MIR),构建重组质粒pET-32a(+)-HBcAg-PEDV S1,转化到感受态细胞BL21(DE3)中,经IPTG诱导表达,SDS-PAGE鉴定纯化后的重组蛋白,通过透射电镜观察其形态结构。结果显示,成功构建重组质粒pET-32a(+)-HBcAg-PEDV S1,并在BL21(DE3)中以包涵体形式表达,纯化、复性后的重组蛋白经过2%磷钨酸负染后透射电子显微镜检测到病毒样颗粒结构。HBcAg-PEDV S1重组蛋白能够自发形成病毒样颗粒,在原核表达中,乙肝核心抗原可作为载体呈现PEDV S1抗原表位,为今后新型PED疫苗的研究提供了思路。A study was undertaken to construct a virus-like particle that uses hepatitis B core antigen(HBcAg)as a carrier to present PEDV S1 epitope.In this work,the 270 bp gene containing the B cell epitope in the PEDV S1 protein was inserted into the main immunodominant region(MIR)of the HBcAg to construct the recombinant plasmid pET-32a(+)-HBcAg-PEDV S1 and transform it into competent cell BL21(DE3),the expression was induced by IPTG,and the expressed protein was purified with Ni column.The purified recombinant protein was identified by SDS-PAGE,and its morphology was detected by transmission electron microscope.The recombinant plasmid pET-32a(+)-HBcAg-PEDV S1 was successfully constructed and expressed as inclusion bodies in BL21(DE3).The purified and renatured recombinant protein,virus-like particle structure was detected by transmission electron microscope after 2%phosphotungstic acid negative staining.HBcAg-PEDV S1 recombinant protein can spontaneously form a virus-like particle structure.In prokaryotic expression,HBcAg can be used as a carrier to present PEDV S1 epitope,which provides ideas for future research on new PED vaccines.

关 键 词：猪流行性腹泻病毒 S1抗原表位 乙肝核心抗原 病毒样颗粒 原核表达 

分 类 号：S852.659.6[农业科学—基础兽医学]