Method for constructing reverse genetic system of red-spotted grouper nervous necrosis virus

The invention belongs to the technical field of virus gene engineering of aquatic animals, and particularly relates to a method for constructing a reverse genetic system of red-spotted grouper nervousnecrosis virus (RGNNV). According to the invention, CMV is used as a promoter to construct an eukaryotic expression plasmid containing an RGNNV genome; the RGNNV is rescued by adopting a method of combining human embryo kidney 293T cells with high transfection efficiency and striped snakehead cells (SSN-1) which efficiently replicate the RGNNV, so that the problem of low transfection efficiency of fish cells is solved. 本发明属于水生动物病毒基因工程技术领域，具体涉及一种构建赤点石斑鱼神经坏死病毒（red-spotted grouper nervous necrosis virus,RGNNV）反向遗传系统的方法。本发明以CMV为启动子构建含RGNNV基因组的真核表达质粒，采用高转染效率的人胚肾293T细胞和对RGNNV高效复制的条纹月鳢细胞（striped snakehead cell,SSN-1）组合的方法来拯救RGNNV，克服了鱼类细胞转染效率低的问题。