Prokaryotic soluble expression method of VP3 gene of South African type 2 foot-and-mouth disease virus

Abstract

The invention discloses a prokaryotic soluble expression method of VP3 gene of a South African type 2 foot-and-mouth disease virus. The method comprises the following steps: according to a gene sequence of the South African type 2 foot-and-mouth disease virus, directly coupling an SUMO solubilization expression tag coding gene to the upstream of a VP3 protein coding gene of FMDV SAT2; introducingsix His tag coding genes into the upstream of SUMO; then, according to escherichia coli codon tropism, optimizing the HisSUMO-SAT2-VP3 gene; recombining the gene to a PUC57 plasmid; taking a HisSUMO-SAT2-VP3-PUC57 plasmid as a template; designing a specific primer for amplifying the South African type 2 foot-and-mouth disease virus VP3 gene; carrying out PCR amplification, and carrying out enzymedigestion on a target fragment and an expression vector by using the same restriction enzyme; cloning a target gene into a pET32a (+) prokaryotic expression vector; constructing a recombinant plasmidpET32a-HisSUMO-VP3; transferring the recombinant plasmid into escherichia coli for expression; and purifying recombinant protein. According to the invention, the pET32a-HisSUMO-VP3 prokaryotic expression vector is successfully constructed, expression in the escherichia coli is achieved, the purified target protein has good reactogenicity, and a foundation is laid for subsequent assembly of a SouthAfrican type 2 foot-and-mouth disease virus subunit vaccine.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a prokaryotic soluble expression method of South African type 2 foot-and-mouth disease virus VP3 gene. Background technique [0002] Foot-and-mouth disease (foot-an-mouth disease, FMD) is caused by foot-and-mouth disease virus (Foot-an-mouth disease, FMDV) infection, can spread in cattle, pigs, sheep, goats and a variety of wild cloven-hoofed animals sick. The disease occurs quickly and has many transmission routes. It has repeatedly broken out in the world, causing huge economic losses to the world's farmers. Foot-and-mouth disease virus is composed of 4 main structural proteins VP1, VP2, VP3 and VP4 and seven non-structural proteins 2A, 2B, 2C, 3A, 3B, 3C, and 3D. There are multiple discontinuous epitopes in the VP3 protein . In countries where FMD is prevalent, the control of the epidemic mainly relies on the existing traditional inactivated vaccines, ...

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Problems solved by technology

However, in the production process of inactivated vaccines, there is a risk of virus inactivation due to incomple

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