Efficient application of a baculovirus-silkworm larvae expression system for obtaining porcine circovirus type 2 virus-like particles for a vaccine
文献类型： 外文期刊
作者： He, Qianhua 1 ; Cao, Zhenming 1 ; Wang, Pengwei 1 ; Lu, Qiuyuan 1 ; Zheng, Hao 1 ; Sun, Jingchen 1 ;
作者机构： 1.South China Agr Univ, Coll Anim Sci, Guangdong Prov Key Lab Agroanim Genom & Mol Breed, Guangzhou 510642, Guangdong, Peoples R China
2.South China Agr Univ, Coll Anim Sci, Subtrop Sericulture & Mulberry Resources Protect, Guangzhou 510642, Guangdong, Peoples R China
期刊名称：ARCHIVES OF VIROLOGY （ 影响因子：2.574； 五年影响因子：2.466 ）
ISSN： 0304-8608
年卷期： 2020 年 165 卷 10 期
页码：
收录情况： SCI
摘要： Porcine circovirus type 2 (PCV2) is a major pathogen associated with swine diseases. It is the smallest single-stranded DNA virus, and its genome contains four major open reading frames (ORFs). ORF2 encodes the major structural protein Cap, which can self-assemble into virus-like particles (VLPs)in vitroand contains the primary antigenic determinants. In this study, we developed a high-efficiency method for obtaining VLPs and optimized the purification conditions. In this method, we expressed the protein Cap with a 6x His tag using baculovirus-infected silkworm larvae as well as theE. coliBL21(DE3) prokaryotic expression system. The PCV2 Cap proteins produced by the silkworm larvae andE. coliBL21(DE3) were purified. Cap proteins purified from silkworm larvae self-assembled into VLPsin vitro, while the Cap proteins purified from bacteria were unable to self-assemble. Transmission electron microscopy confirmed the self-assembly of VLPs. The immunogenicity of the VLPs produced using the baculovirus system was demonstrated using an enzyme-linked immunosorbent assay (ELISA). Furthermore, the purification process was optimized. The results demonstrated that the expression system using baculovirus-infected silkworm larvae is a good choice for obtaining VLPs of PCV2 and has potential for the development of a low-cost and efficient vaccine.
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