Construction and identification of recombinant feline herpesvirus expressing VP1 protein of feline calicivirus
10.16303/j.cnki.1005-4545.2025.08.06
- VernacularTitle:表达猫杯状病毒VP1蛋白的重组猫疱疹病毒的构建与鉴定
- Author:
Lisi AI
1
;
Cuicui JIAO
;
Hongli JIN
;
Pei HUANG
;
Haili ZHANG
;
Yuanyuan LI
;
Hualei WANG
Author Information
1. 吉林大学动物医学学院/人畜共患病研究所人畜共患传染病重症诊治全国重点实验室/人畜共患病研究教育部重点实验室,吉林长春 130062
- Publication Type:Journal Article
- Keywords:
feline calicivirus;
feline herpesvirus type Ⅰ;
VP1 protein of feline calicivirus;
recombinant feline herpesvirus;
biological characteristics
- From:
Chinese Journal of Veterinary Science
2025;45(8):1624-1631,1641
- CountryChina
- Language:Chinese
-
Abstract:
Feline herpesvirus type Ⅰ(FHV-1)was used as the vector.The gI and gE genes of FHV-1 were replaced with the feline calicivirus(FCV)VP1 gene and the red fluorescent protein(mCherry)gene by CRISPR/Cas9 systems and homologous recombination technology,and the re-combinant virus strain FHV △gI&gE/VP1-mCherry+was successfully rescued.The recombinant virus strain was purified by plaque assay.The biological characteristics and genetic stability of the recombinant virus were analyzed by indirect immunofluorescence assay,plaque morphological anal-ysis,and PCR.The results of the indirect immunofluorescence identification showed that the re-combinant virus FHV △gI&gE/VP1-mCherry+could express the VP1 protein in F81 cells,and the growth characteristics of the recombinant virus were not significantly different from those of the parent virus FHV-1.The plaque morphology and staining results indicated that the area of the plaque formed by the recombinant virus was smaller than that of the parent virus,suggesting that the spread ability of the recombinant virus between cells was reduced after the deletion of the gI and gE genes.The result of PCR showed that the VP1 gene could still be detected after 15 succes-sive passages of the recombinant virus,indicating that the recombinant virus had good genetic stability.In this study,the recombinant virus strain expressing the FCV VP1 protein was successfully prepared,which will lay a foundation for the development of engineered FCV and FHV-1 vaccine.
