A method for constructing a reverse genetic system of red-spotted grouper neuronecrosis virus

A technology of red-spotted grouper and nerve necrosis, applied in the field of aquatic animal virus genetic engineering, can solve the problem of low transfection efficiency and achieve the effect of convenient virus rescue
CN111471710BActive Publication Date: 2022-03-08HUAZHONG AGRI UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
HUAZHONG AGRI UNIV
Publication Date
2022-03-08

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Abstract

The invention belongs to the technical field of aquatic animal virus genetic engineering, and in particular relates to a method for constructing a red-spotted grouper nervous necrosis virus (RGNNV) reverse genetics system. The present invention uses CMV as a promoter to construct a eukaryotic expression plasmid containing the RGNNV genome, and adopts a method of combining human embryonic kidney 293T cells with high transfection efficiency and striped snakehead cells (SSN‑1) that can efficiently replicate RGNNV To rescue RGNNV, it overcomes the problem of low transfection efficiency of fish cells.
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Description

technical field

[0001] The invention belongs to the technical field of aquatic animal virus genetic engineering, and specifically relates to a method for constructing a reverse genetic system of red grouper neuronecrosis virus. Background technique

[0002] Nervous necrosis virus (NNV) is a member of the Nodaviridae family. The disease caused by its infection is called viral nerve necrosis (viral nervous necrosis, VNN) or viral encephalopathy and retinopathy (viral encephalopathy and retinopathy, VER). It has now been reported in more than 120 different farmed and wild fish species, causing huge economic losses to the aquaculture industry. In 2001, Lin Li et al. isolated the virus from naturally occurring red-spotted grouper fry in Daya Bay, Guangdong Province, China, and it was red-spotted grouper nervous necrosis virus (RGNNV). RGNNV is a positive-strand RNA virus, and the viral genome contains two segments of RNA (RNA1 and RNA2). RNA1 encodes viral RNA polymerase, and ...

Claims

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