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

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。

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

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.