Construction and identification of recombinant fowl adenovirus 4 expressing Cap protein of goose astrovirus virus genotype 2
10.16303/j.cnki.1005-4545.2025.03.07
- VernacularTitle:表达GoAstV-2 Cap蛋白的重组FAdV-4的构建与鉴定
- Author:
Xingyu LI
1
;
Yan LI
;
Panpan YANG
;
Junjie LIU
;
Mengjia XIANG
;
Yutao ZHU
;
Luyao QIU
;
Qilong QIAO
;
Boshun ZHANG
;
Dexin BU
;
Chenghao HAN
;
Chunmei YU
;
Yanfang CONG
;
Zeng WANG
;
Jianli LI
;
Baiyu WANG
;
Jun ZHAO
Author Information
1. 河南农业大学 动物医学院,河南 郑州 450046
- Publication Type:Journal Article
- Keywords:
goose astrovirus genotype 2;
fowl adenovirus 4;
Cap protein;
inactivated bivalent vaccine
- From:
Chinese Journal of Veterinary Science
2025;45(3):443-448,513
- CountryChina
- Language:Chinese
-
Abstract:
To construct a recombinant fowl adenovirus 4(FAdV-4)expressing the Cap protein of goose astrovirus genotype 2(GoAstV-2),the expression cassette of Cap gene was inserted into the natural 1 966 bp deletion region of the FAdV-4 genome in the infectious clone p15A-cm-FAdV4-HNJZ.The resulted recombinant plasmid p15A-cm-FAdV4-HNJZ-Cap/GoAstV-2 was linearized with restriction enzyme and transfected into chicken hepatoma cell line(LMH)to rescue the recombinant FAdV-4 expressing the Cap protein of GoAstV-2,rF Ad V4-Cap/GoAstV-2.After 15 passages in LMH cells,the recombinant rFAdV4-Cap/GoAstV-2 was identified by PCR using primers flanking the insertion site of the Cap gene expression cassette and using viral genome DNA extracted from rFAdV4-Cap/GoAstV-2 infected LMH cells as template.LMH cells were in-fected with 15th passage rFAdV4-Cap/GoAstV-2 and indirect immunofluorescence was performed with a polyclonal antibody against Cap protein as the primary antibody.Western blot was carried out with lysates of rFAdV4-Cap/GoAstV-2 infected LMH cells.The in vitro replication dynamic of the 15th passage of the rFAdV4-Cap/GoAstV-2 was also investigated in LMH cells.The results demonstrated that the Cap gene of GoAstV-2 was presented in the genome of the recombinant vi-rus rF AdV4-Cap/Go Ast V-2,and could be expressed stably.The prepared recombinant virus in this study will lay a foundation for developing inactivated bivalent vaccine candidate against co-in-fection of FAdV-4 and GoAstV-2 in goose.
