Effect of PK-15 cell line stably expressing UL54 protein on replication efficiency of UL54 gene deletion recombinant pseudo rabies virus
10.16303/j.cnki.1005-4545.2025.06.03
- VernacularTitle:稳定表达UL54蛋白的PK-15细胞株对UL54基因缺失重组伪狂犬病病毒复制效率的影响
- Author:
Mingzhi LI
1
;
Hongxia WU
;
Yimin WANG
;
Yuan SUN
Author Information
1. 河南科技学院动物科技学院,河南新乡 453003;中国农业科学院哈尔滨兽医研究所动物疫病防控全国重点实验室,黑龙江哈尔滨 150069
- Publication Type:Journal Article
- Keywords:
PRV;
lentivial vector;
UL54 gene;
PK-15 cell
- From:
Chinese Journal of Veterinary Science
2025;45(6):1117-1125
- CountryChina
- Language:Chinese
-
Abstract:
The UL54 protein is a nonstructural protein of the pseudorabies virus(PRV).Electron microscopy observations showed that most of particles of recombinant PRV(rPRV-△UL54)with a deletion of the UL54 gene had only a nucleocapsid and lacked an intact vesicle structure.The in-complete structure of the viral particles led to a significant reduction in the infection efficiency of the virus,which could not be applied in the study of the gene function and mechanism of action of UL54.In this study,the UL54 gene was cloned into the lentiviral vector pLVX-IRES-Puro,and af-ter lentiviral transduction,the UL54 gene was integrated into the PK-15 cell genome.A cell line(PK-15-UL54)overexpressing UL54 protein was successfully constructed and characterized by PCR,indirect immunofluorescence assay and protein immunoblotting.Replication was assayed in the PK-15-UL54 cell line after inoculation with UL54 gene deletion virus(rPRV-△UL54-Re).Elec-tron microscopy observation showed that the rPRV-△UL54-Re virus had a complete viral particle structure.Fluorescence microscopy observations showed that rPRV-△UL54-Re virus could normal-ly infect cells and replicate.The results of quantitative PCR assay showed that the replication efficiency of rPRV-△UL54-Re virus increased.Therefore,PK-15-UL54 cell line is an important tool to improve the replication efficiency of UL54 gene deletion recombinant pseudorabies virus and plays an important role in exploring the mechanistic study of UL54 gene regulation of PRV repli-cation.
