表达猪圆环病毒3型Cap蛋白重组伪狂犬病病毒的构建及其对小鼠免疫效果评价

doi:10.11843/j.issn.0366-6964.2025.11.035

Abstract

Abstract:

This study aimed to construct a viral vector vaccine expressing the Cap protein of porcine circovirus type 3 (PCV3). In this study, pseudorabies virus (PRV) SX1910 strain was designated as the parental strain. Using CRISPR/Cas9 and homologous recombination technologies, a recombinant virus SX1910-ΔgE/gI-GFP was constructed, which lacked the gE and gI genes and harbored a LoxP site and GFP fluorescent marker. Then, the GFP expression cassette of SX1910-ΔgE/gI-GFP was replaced with the double expression cassette of the PCV3 Cap gene by using Cre enzyme, generating the recombinant virus rSX1910-ΔgE/gI-2Cap_PCV3. Then its in vitro biological characteristics and immunological effects were analysed. The results showed that rSX1910-ΔgE/gI-2Cap_PCV3 exhibited significantly reduced in vitro replication capacity and plaque size compared to the parental strain SX1910. Preliminary evaluation of immunogenicity in 6-week-old female BALB/c mice demonstrated that rSX1910-ΔgE/gI-2Cap_PCV3 effectively stimulated the production of Cap protein-specific antibodies in mice, alleviated PCV3-induced lung tissue damage, and significantly reduced viral loads in tissues. Therefore, rSX1910-ΔgE/gI-2Cap_PCV3 can elicit a good immune response in mice, providing a scientific foundation for the development of novel vaccines for PCV3.

Key words: porcine circovirus type 3, Cap protein, pseudorabies virus, CRISPR Cas9, immune protection

CLC Number:

S852.659.1

SONG Ruoqi, TAN Shanshan, MA Ruiyi, CHEN Xinxin, YI Yue, MA Mengyao, WANG Ying, NIU Sheng, YAN Fang, ZHAO Yujun, TIAN Wenxia, REN Jianle. Construction and Immune Effect in Mice of Recombinant Pseudorabies Virus Expressing Porcine Circovirus Type 3 Cap Protein[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(11): 5778-5788.

Figures/Tables 7

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引物名称 Primer name	序列(5′→3′) Sequence (5′→3′)	酶切位点	产物大小/bp Product/bp	用途 purpose
PCV3-3.1-F/R	F: CTAGCGTTTAAACTTAAGCTTATGAGACACAGA GCTATATT	Hind Ⅲ	687	扩增Cap基因
PCV3-3.1-F/R	R: AACGGGCCCTCTAGACTCGAGTTAGAGAACG GACTTGTAAC	XhoⅠ	687	扩增Cap基因
Cap3-SK1-F/R	F: CTATAGGGCGAATTGGGTACCGACATTGATTA TTGACTAGT	KpnⅠ	1 637	扩增Cap表达盒1
Cap3-SK1-F/R	R: TCCCCCGGGCTGCAGGAATTCCCATAGAGC CCACCGCATCC	EcoRⅠ	1 637	扩增Cap表达盒1
Cap3-SK2-F/R	F: TTCCTGCAGCCCGGGGGATCCGACATTGATTA TTGACTAGT	BamHⅠ	1 639	扩增Cap表达盒2
Cap3-SK2-F/R	R: AGCTCCACCGCGGTGGCGGCCGCCCATAGAG CCCACCGCATCC	Not Ⅰ	1 639	扩增Cap表达盒2
2Cap_PCV3-F/R	F: ATAACTTCGTATAGCATACATTATACGAAGTTATC TCACTATAGGGCGAATTGGGTACCGACATTGA		3 262	扩增和鉴定双拷贝的Cap基因表达
2Cap_PCV3-F/R	R: ATAACTTCGTATAATGTATGCTATACGAAGTTAT GAGCTCCACCGCGGTGGCGGCCGCCCATAGAG		3 262	扩增和鉴定双拷贝的Cap基因表达
QPCV3-F/R	F: CACGGAGGTCTGTAGGGAGA R: GGACACTCGTAGCACCACAA		156	荧光定量

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Construction and Immune Effect in Mice of Recombinant Pseudorabies Virus Expressing Porcine Circovirus Type 3 Cap Protein

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