Preparation method of grass carp reovirus genetic engineering vaccine

Abstract

The invention provides a method for preparing grass carp reovirus (GCRV) gene engineering vaccines. The method comprises the following steps of: selecting recombinant baculoviruses (vAcGCRV-VP5 / VP7) of sf9 insect cell proliferated recombinant GCRV outer capsid proteins VP5 and VP7; separating recombinant protein components by freezing, thawing and cracking recombinant virus infected sf9 cells and adopting sucrose lining centrifuging method, and thus obtaining a purified in vitro expression VP5 and VP7 protein complex; and performing fish protection experiments on the purified VP5 and VP7 proteins to prove that the VP5 and VP7 proteins have good immune protection effect. The GCRV gene engineering vaccines prepared by the method not only have good stability and immunogenicity, but also have good immune protection effect on grass carp fries; the immune protection rate of the GCRV gene engineering vaccines reaches 90 percent; and the method is suitable for large-scale production and application of the vaccines.

Description

technical field [0001] The invention relates to the field of genetic engineering / molecular virology, in particular to the preparation of a genetically engineered vaccine for the outer capsid protein of Grass carp reovirus (GCRV, also known as Grass carp hemorrhagic virus, GCHV) method. The virus strain used in the present invention is grass carp reovirus Hunan Shaoyang strain GCRV873, also known as grass carp haemorrhagic disease virus Hunan Shaoyang strain GCHV873, has been preserved by the China Center for Type Culture Collection, the preservation date is July 23, 2004, and the preservation number is CCTCC NO: V200414. Background technique [0002] grass carp( Ctenopharyngodon idellus ) is one of the four major domestic fishes in my country's freshwater aquaculture. It is cheap and good, and is deeply loved by consumers. Grass carp reovirus is the main pathogen that causes the death of grass carp. When the disease is severe, it can cause more than 85% of grass carp fry ...

AI Technical Summary

Problems solved by technology

[0003] In view of the instability of crude tissue plasma, cell inactivated vaccines and antigen subunit vaccines and genetic material such as residual nucleic acids, as well as the high cost and unstable potency of virus particle subunit vaccines obtained by conventional virus purification methods, etc. , it is necessary to improve the previous preparation method