家畜生态学报2026，Vol.47Issue(8)：88-94,7.DOI:10.3969/j.issn.1673-1182.2026.08.011
猪戊型肝炎病毒ORF2蛋白在杆状病毒系统中的表达与纯化
Expression and Purification of the Porcine Hepatitis E Virus ORF2 Protein Using the Baculovirus Expression System
摘要
Abstract
This study aimed to produce highly active ORF2 protein of porcine hepatitis E virus(HEV),optimize the preparation strategy for virus-like particle(VLP)vaccines,and provide a new approach for the development of low-cost,high-yield vaccines against porcine HEV.Using a plasmid containing the ORF2 gene sequence of a porcine HEV strain as the template,the recombinant bacmid rBac-SDS-HEV-p495 carrying the ORF2 gene was constructed by RT-PCR amplification,restriction enzyme digestion,and ligation.The recombinant bacmid was then transfected into Spodoptera frugiperda 9(Sf9)cells to generate recombinant baculovirus.Sf9 cells were infected with the recombinant baculovirus,and expression of the target protein was confirmed by Western blotting and indirect immunofluorescence assay(IFA).The ex-pressed ORF2 protein was subsequently purified by sucrose density gradient centrifugation and ion-ex-change chromatography.The results showed that the recombinant baculovirus rBac-SDS-HEV-p495 was successfully constructed.Following infection of Sf9 cells,the porcine HEV ORF2 protein with an apparent molecular weight of approximately 55 kDa was successfully expressed in both cell lysates and culture su-pernatants.The expression level of the target protein in the culture supernatant reached 20 mg/L.In con-clusion,a highly active porcine HEV ORF2 protein was successfully expressed using the baculovirus ex-pression system,providing an important foundation for the development of novel vaccines and diagnostic reagents against porcine HEV.关键词
猪戊型肝炎病毒/重组杆状病毒/SF9细胞/蛋白纯化Key words
porcine hepatitis E virus/recombinant baculovirus/SF9 cell/protein purification分类
农业科技引用本文复制引用
张园..猪戊型肝炎病毒ORF2蛋白在杆状病毒系统中的表达与纯化[J].家畜生态学报,2026,47(8):88-94,7.基金项目
陕西省教育厅专项科研计划项目(24JK0669) （24JK0669）
西安职业技术学院博士科研启动项目(2025QDJF04) （2025QDJF04）
创新能力支撑计划-陕西省青年科技新星项目(2026RC-KJXX-082) （2026RC-KJXX-082）
