Scientia Agricultura Sinica ›› 2015, Vol. 48 ›› Issue (5): 976-986.doi: 10.3864/j.issn.0578-1752.2015.05.16

• ANIMAL SCIENCE·VETERINARY SCIENCERE • Previous Articles     Next Articles

PCV2 Virus Like Particles Vaccine Produced with Recombinant Cap Protein Expressed in E.coli

ZHAO Xiao-yun1,2, QIAO Xu-wen1, CHEN Jin1, LI Peng-cheng1, YU Xiao-ming1, ZHU Guo-qiang2, ZHENG Qi-sheng1, HOU Ji-bo1   

  1. 1National Research Center of Veterinary Biological Engineering and Technology, Nanjing 210014
    2College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, Jiangsu
  • Received:2013-12-26 Online:2015-03-01 Published:2015-03-01

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Abstract: 【Objective】The objective of this study is to develop recombinant virus like particles vaccine for Porcine circovirus type 2 through soluble prokaryotic expression of ORF2 gene. 【Method】 The optimized ORF2 gene of PCV2 NJ strain was synthesized according to codon usage of E.coli, and then the optimized ORF2 gene was cloned into pQZ1 to get the recombinant prokaryotic expression vector named pQZ-Cap. The target gene was expressed with 1.0 mmol·L-1 IPTG induction for 24 h under 15℃ on the prokaryotic expression platform in authors’ laboratory following the recombinant plasmid pQZ-Cap transformed into host E.coli BL21. Firstly, SDS-PAGE and Western blotting were used to identify the expression and solubility of the recombinant protein. Secondly, assemble for recombinant Cap VLP was evaluated through electron microscope analysis technology. Furthermore, swines were inoculated with recombinant Cap protein emulisified in SPPEIC 206 adjuvant, and the immunogenicity of recombinant Cap protein vaccine was evaluated by PCV2 specific antibody detection and virus attack.【Result】SDS-PAGE and Western blotting results indicated that the optimized ORF2 gene of PCV2 NJ strain could be efficiently expressed in E.coli BL21 in form of completely soluble. Abundant PCV2 VLP with diameter about 17 nm could be observed through electron microscope, exist in supernatant of the induced E.coli after ultrasonication. VLP vaccine composed of recombinant Cap protein possessed satisfactory immunity. Swine immunized with this VLP vaccine generated perfect antibody response with 100% qualified at 21 d post one sole vaccination and could be completely protected from pathogenic virus attack.【Conclusion】To summarize, optimized ORF2 gene of PCV2 NJ strain was successfully expressed in E.coli completely soluble and recombinant Cap protein can auto-assemble into VLP particles after ultrasonication. The VLP particles pose perfect immunity and could be used as subunit vaccines to prevent PCV2 infection.

Key words: porcine circovirus type 2, ORF2 gene, soluble expression, VLP particles, immunogenicity

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