Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/93091
標題: Expression and Structural Analysis of the Porcine Circovirus Type 2 Cap Protein
豬環狀病毒第二型Cap蛋白之表現及結構分析
作者: 陳姿伃
Tzu-Yu Chen
關鍵字: 豬環狀病毒第二型
外殼蛋白
蛋白表現
結構
類病毒顆粒
PCV2 Cap
Expression of Cap
VLP
fusion tag
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摘要: Porcine circovirus type 2 (PCV2) is the primary causative agent of the economically important global disease, porcine circovirus associated diseases (PCVAD). The viral capsid protein (Cap) containing the major antigenic sites has become the target protein for developing of diagnostic reagents and PCV2 subunit vaccine. The purpose of this study is to express a variety of recombinant Cap proteins using an E. coli expression system for further analysis of the structure and immunogenicity of viral capsid. Four recombinant Cap proteins including the rCapT7+ containing a N-terminal T7 tag, the rCapHis+ containing a C-terminal His tag, the rCapT7+His+ containing both tags at each end, and the rCap containing no tag were constructed. The recombinant Cap proteins were purified by the continuous sucrose gradient ultracentrifugation and confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. All the expressed Cap proteins no matter containing a fusion tag could self-assemble into virus-like particles (VLPs) by the transmission electronmicroscope analysis. The immunogenicity of PCV2 VLPs were further evaluated in mice. Both the rCap and fusion tags containing rCapT7+His+ VLPs immunized groups were able to elicit specific antibody responses to PCV2 with neutralizing antibody titers of 1:8 after twice booster immunization. In addition, both immunized groups showed detectable of IFN- secreting cells response in the ELISpot analysis. All the results demonstrated that the PCV2 VLPs based on the E. coli-expressed Cap proteins could mount both humoral and cellular immune response against PCV2 Cap in mice, and the terminal fusion tags had no significant effect on structural assembly and immunogenicity of Cap protein.
URI: http://hdl.handle.net/11455/93091
文章公開時間: 2018-06-22
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