Immunogenicity of recombinant porcine pseudorabies virus expressing ORF2 gene of porcine circovirus type 2d
10.16303/j.cnki.1005-4545.2025.10.04
- VernacularTitle:猪圆环病毒2d基因型ORF2基因重组猪伪狂犬病病毒的免疫原性
- Author:
Xianqin JIAO
1
;
Tao WANG
;
Runbo TIAN
;
Shijie MA
;
Zhihao YAN
;
Hongying CHEN
Author Information
1. 河南农业大学动物医学院,河南 郑州 450046
- Publication Type:Journal Article
- Keywords:
porcine circovirus type 2d genotype;
porcine pseudorabies virus;
live vector vaccine;
ORF2 gene
- From:
Chinese Journal of Veterinary Science
2025;45(10):2118-2125
- CountryChina
- Language:Chinese
-
Abstract:
To develop vaccines for the prevention and control of porcine circovirus 2d genotype(PCV2d)and pseudorabies virus(PRV),the PCV2d ORF2 gene was amplified by PCR,and cloned into the BamH Ⅰ site of PRV transfer plasmid pG vector harboring the enhanced green fluorescent protein(EGFP)gene.The resulting recombinant transfer plasmid pG-PCV2d-EGFP was transfect-ed into ST cells infected with the three gene deleted PRV variant strain gE-/g-/TK-PRV NY to generate a recombinant virus rPRV-PCV2d-EGFP+,and then the EGFP gene was knocked out to harvest the rPRV-PCV2d using gene-editing technology termed CRISPR/Cas9 system.The recom-binant virus rPRV-PCV2d had similar genetic stability to the parental PRV as indicated by PCR and one-step growth curve test,and the expression of PCV2d capsid(Cap)protein was validated by Western blot.In animal experiment,higher PCV2-specific ELISA antibodies and detectable PCV2-specific neutralizing antibodies could be elicited in mice immunized with the recombinant vi-rus rPRV-PCV2d compared to commercial PCV2 inactivated vaccine.rPRV-PCV2d significantly re-duced the PCV2d loads in tissues such as the heart,liver and spleen of mice following virulent PCV2d challenge.Moreover,rPRV-PCV2d elicits PRV-specific immune responses in mice and can prevent PRV virulent infection in mice,indicating the recombinant virus rPRV-PCV2d has strong immunogenicity.
