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AssigneeYangzhou University
InventorsNanhua Chen, Ming Qiu, Hong Lin, Jianzhong Zhu
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Disclosed are an rBJ-VVL plasmid, a mutant strain of NADC34-like PRRSV-2 and a preparation method therefor and application thereof. Further disclosed is an NADC34-like PRRSV-2-specific vaccine. In the present disclosure, a modified strain rBJ-VVL with tropism for Marc-145 cells is obtained by precisely mutating an amino acid at positions 91/97/98 of GP2a; the modified virus constructed in the present disclosure can be propagated in Marc-145 cells, cause cytopathic effects and form plaques when inoculated into Marc-145 cells for serial passage; the resulting Marc-145 cell-passaged viruses have an extremely viral load, and a large number of new progeny viruses can be obtained in a short time; and the Marc-145-adapative modified strain cultured in the present disclosure is used to create a first NADC34-like PRRSV-2-specific vaccine.
An rBJ-VVL plasmid, wherein the rBJ-VVL plasmid is obtained by using a plasmid of a reverse genetics platform pACYC177-rBJ1805-2 as a template, designing a primer pair for amino acid mutations at positions 91, 97, and 98 of GP2a protein of an rBJ1805-2 virus to V, V and L, respectively, amplifying to obtain a mutated fragment by a PCR amplification method and then ligating the mutated fragment with a linearized vector;
A mutant strain of NADC34-like PRRSV-2, wherein the mutant strain is obtained by mutating amino acids at positions 91, 97, and 98 of a GP2a protein of an infectious clone virus rBJ1805-2 of NADC34-like PRRSV-2 from T, M and F to V, V and L, respectively;
A method for constructing an rBJ-VVL plasmid, comprising the following steps:
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