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

doi:10.11843/j.issn.0366-6964.2021.06.031

畜牧兽医学报 ›› 2021, Vol. 52 ›› Issue (6): 1760-1764.doi: 10.11843/j.issn.0366-6964.2021.06.031

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

刘如月¹, 范京惠¹, 刘涛², 苑军辉³, 李清艳¹, 翟向和^1*, 左玉柱^1*

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

收稿日期:2020-11-09 出版日期:2021-06-23 发布日期:2021-06-22
通讯作者: 左玉柱,主要从事动物传染病学的教学、科研和社会服务,E-mail:zuoyuzhu@163.com;翟向和,主要从事基础兽医学的教学、科研工作,E-mail:ZXH958@163.com
作者简介:刘如月(1996-),女,河北衡水人,硕士生,主要从事动物传染病防控研究,E-mail:2371604609@qq.com
基金资助:
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）

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

LIU Ruyue¹, FAN Jinghui¹, LIU Tao², YUAN Junhui³, LI Qingyan¹, ZHAI Xianghe^1*, ZUO Yuzhu^1*

1. College of Veterinary Medicine/Agriculture Science and Education Center, Hebei Agricultural University, Baoding 071001, China;
2. Ringpu(Baoding) Biological Pharmaceutical Co. Ltd., Baoding 071001, China;
3. Xingtang Animal Health Supervision Institution, Xingtang 050600, China

Received:2020-11-09 Online:2021-06-23 Published:2021-06-22

摘要/Abstract

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。

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

Abstract: 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.

Key words: porcine epidemic diarrhea virus, S1 epitope, hepatitis B core antigen, virus-like particles, prokaryotic expression

中图分类号:

S852.659.6

刘如月, 范京惠, 刘涛, 苑军辉, 李清艳, 翟向和, 左玉柱. 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[J]. 畜牧兽医学报, 2021, 52(6): 1760-1764.

LIU Ruyue, FAN Jinghui, LIU Tao, YUAN Junhui, LI Qingyan, ZHAI Xianghe, ZUO Yuzhu. Construction and Identification of Hepatitis B Core Antigen Virus-like Particles Carrying PEDV S1 Epitope[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(6): 1760-1764.

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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

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