Construction and identification of foot-and-mouth disease virus replicon system carrying fluorescent protein UnaG
10.16303/j.cnki.1005-4545.2025.07.08
- VernacularTitle:携带荧光蛋白UnaG的口蹄疫病毒复制子系统的构建及其鉴定
- Author:
Xuetong LI
1
;
Yuanyuan HUANG
1
;
Xianglong GUO
1
;
Yun ZHANG
1
;
Shiqi SUN
1
;
Huichen GUO
1
;
Jinen WU
1
Author Information
1. 中国农业科学院兰州兽医研究所 兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室,甘肃兰州 730000
- Publication Type:Journal Article
- Keywords:
UnaG;
foot-and-mouth disease virus;
replicator
- From:
Chinese Journal of Veterinary Science
2025;45(7):1411-1416,1436
- CountryChina
- Language:Chinese
-
Abstract:
To construct a subgenomic replication subsystem of foot-and-mouth disease virus(FM-DV)carrying fluorescent protein(UnaG),the full-length cDNA plasmid of FMDV was employed as the template.A few of structural and non-structural protein genes of FMDV were removed by double-enzyme digestion.By substituting reporter gene sequences expressing the green fluorescent protein UnaG for specific structural protein sequences of FMDV,FMDV-UnaG replicators were successfully created.After PCR and sequencing,linearized FMDV-UnaG replicons were transfected into BSR/T7 cells expressing T7 RNA polymerase,and the fluorescence signal was observed through fluorescence microscopy and laser confocal technique.The results demonstrated that the constructed FMDV-UnaG replicons could effectively express UnaG protein,and the protein colo-calized with FMDV 3A protein.Additionally,Western blot and RT-qPCR also detected that the replicator RNA could express the non-structural proteins of the virus and replicate autonomously in BSR/T7 cells,respectively.In conclusion,the successful construction of FMDV-UnaG sub-genomic replicators offers a favorable tool for further research on the replication and translation mechanism of FMDV and the development of vaccine vectors.
