Construction of stable BHK-21 cell lines overexpressing APN of different species and the susceptibility to different coronaviruses
10.16303/j.cnki.1005-4545.2025.10.01
- VernacularTitle:过表达APN稳转BHK-21细胞系的构建及其对冠状病毒的易感性
- Author:
Dan WANG
1
;
Hengjie ZHANG
;
Yuyang TIAN
;
Xiaohan HOU
;
Zeao CHEN
;
Ying HU
;
Wenchao ZHANG
;
Jianle REN
;
Ying WANG
;
Yujun ZHAO
;
Ding ZHANG
;
Bo YANG
;
Wenxia TIAN
;
Sheng NIU
Author Information
1. 山西农业大学动物医学学院,山西 晋中 030801
- Publication Type:Journal Article
- Keywords:
APN;
BHK-21 cell lines;
cross-species transmission;
porcine deltacoronavirus;
transmis-sible gastroenteritis virus
- From:
Chinese Journal of Veterinary Science
2025;45(10):2095-2101
- CountryChina
- Language:Chinese
-
Abstract:
This study aims to establish BHK-21 stable cell lines expressing APN from four species(human,pig,dog,and cat),the APN fragments were amplified from pEGFP-C1-APN plasmids of the four species stored in the laboratory to generate the recombinant plasmids pcDNA4.0-APN.Af-ter the recombinant plasmids were transfected into BHK-21 cells,the stable BHK-21 cell lines ex-pressing the APNs were selected by two rounds of limited dilution.The constructed BHK-21 cell lines were identified by indirect immunofluorescence assay(IFA),and their susceptibility to PD-CoV and TGEV was tested for these four cell lines.Virus infection experiments revealed that PD-CoV infected cells expressing human,pig,and dog APNs,while it did not infect cells expressing cat APN.Simultaneously,TGEV infected cells expressing pig,dog,and cat APNs,but did not infect cells expressing human APN.The results suggest that the risk of cross-species infection for different coronaviruses and the established cell line can be used effectively to evaluate the virus in-fection.The findings also revealed that PDCoV has the potential risk of cross-species infection of human and dog,and TGEV has the potential risk of cross-species infection of dog and cat.These results provide a basis for the prevention and control strategy of coronaviruses.
