Method for preparing grass carp reovirus (GCRV) gene engineering vaccines
A technology of genetic engineering vaccine and reovirus, applied in the field of genetic engineering/molecular virology, can solve the problems of high cost and unstable titer of virus particle subunit vaccine, improve the immune protection rate of fish, good stability Sexual and immunogenic, high expression effect
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[0034] The grass carp reovirus genetically engineered vaccine provided by the invention is a genetically engineered vaccine that expresses GCRVVP5 and VP7 efficiently in insect cells obtained by a genetic engineering method. The specific preparation steps of the GCRV genetic engineering vaccine are as follows: through sucrose-lined centrifugation, the purified VP5 and VP7 protein complexes are obtained, the purified protein is diluted with 0.65% normal saline and then soaked in fry to obtain an immune protection effect of 90%.
[0035] 1. Propagate the recombinant baculoviruses of the recombined GCRV outer capsid proteins VP5 and VP7 using the sf9 insect cell line, collect the sf9 cells infected with the recombinant virus for 72 hours, centrifuge the cells at a low speed, and remove the supernatant. Rinse 3 times with PBS or phosphate buffered saline to remove bovine serum albumin components, add 3 times the volume of lysate to the low-speed centrifuged cell pellet, mix thoroug...
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