猪圆环病毒3型Cap重组鞭毛蛋白融合表达及其免疫原性研究

doi:10.16431/j.cnki.1671-7236.2023.05.031

Abstract

Abstract: 【Objective】 The aim of this experiment was to express the recombinant Cap-Flagellin protein of Porcine circovirus type 3 (PCV3) by Baculovirus expression system and analyze its reactogenicity and immunological properties,providing important support for the development of PCV3 subunit vaccine.【Method】 The Cap gene was optimized by optimal codon and fused with Flagellin gene of Salmonella Typhimurium then cloned into pFastBac^TMⅠ vector.Recombinant Cap-Flagellin Baculovirus was rescued through the Bac-to-Bac system and identified by SDS-PAGE,indirect immunofluorescence assay (IFA) and Western blotting,and then mice were immunized with recombinant Cap-Flagellin Baculovirus to evaluate its immunogenicity.【Result】 Recombinant Cap-Flagellin Baculovirus could effectively express recombinant proteins in Sf9 cells,the target protein bands of 68 ku were detected by SDS-PAGE.IFA results showed that the specific green fluorescence occurred in the membrane and cytoplasm of Sf9 cells 72 h after recombinant Baculovirus infection.Western blotting showed that the recombinant protein could react specifically with PCV3 positive serum,all indicated it had good reactogenicity.The serum Cap-specific antibody levels of mice immunized with Cap-Flagellin Baculovirus on the 21th and 28th day were higher compared with the Cap Baculovirus group(P＜0.05 or P＜0.01),and IL-8 cytokine level of spleen cells in recombinant Cap-Flagellin group was significantly or extremely significantly higher than that of other groups(P＜0.05 or P＜0.01),which indicated that the recombinant Cap-Flagellin protein had better immunogenicity.【Conclusion】 In this study,PCV3 Cap recombinant flagellin was successfully expressed by Baculovirus expression system and had good biological activity which laid a good foundation for the development of PCV3 subunit vaccine.

Key words: Porcine circovirus 3 (PCV3); recombinant Baculovirus; Cap protein; flagellin; immune property

CLC Number:

S852.65^＋1

JIANG Haijun, WANG Dan, GUO Yu, WEI Li, HOU Lei, WANG Jing, ZHU Shanshan, LIU Jue. Fusion Expression and Immunogenicity of Recombinant Cap-flagellin Protein of Porcine Circovirus Type 3[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2044-2052.

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Fusion Expression and Immunogenicity of Recombinant Cap-flagellin Protein of Porcine Circovirus Type 3

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