Construction of Attenuated Strains for Red-Spotted Grouper Nervous Necrosis Virus (RGNNV) via Reverse Genetic System
Yingying Lei,
Yu Xiong,
Dagang Tao,
Tao Wang,
Tianlun Chen,
Xufei Du,
Gang Cao,
Jiagang Tu,
Jinxia Dai
Affiliations
Yingying Lei
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China
Yu Xiong
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China
Dagang Tao
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China
Tao Wang
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China
Tianlun Chen
Institute of Neuroscience, State Key Laboratory of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Chinese Academy of Sciences, Shanghai 200031, China
Xufei Du
Institute of Neuroscience, State Key Laboratory of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Chinese Academy of Sciences, Shanghai 200031, China
Gang Cao
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China
Jiagang Tu
College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China
Jinxia Dai
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China
The nervous necrosis virus (NNV) mainly attacks the central nervous system of fish to cause viral nervous necrosis, which is an acute and serious prevalent disease in fish. Among different genotypes of NNV, red-spotted grouper nervous necrosis virus (RGNNV) is the most widely reported, with the highest number of susceptible species. To better understand the pathogenicity of RGNNV, we first developed a reverse genetic system for recombinant RGNNV rescue using B7GG and striped snakehead (SSN-1) cells. Furthermore, we constructed attenuated RGNNV strains rRGNNV-B2-M1 and rRGNNV-B2-M2 with the loss of B2 protein expression, which grew slower and induced less Mx1 expression than that of wild-type RGNNV. Moreover, rRGNNV-B2-M1 and rRGNNV-B2-M2 were less virulent than the wild-type RGNNV. Our study provides a potential tool for further research on the viral protein function, virulence pathogenesis, and vaccine development of RGNNV, which is also a template for the rescue of other fish viruses.
