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, ...

AI Technical Summary

Problems solved by technology

However, in the production process of inactivated vaccines, there is a risk of virus inactivation due to incomplete virus inactivation, which prompts people to develop a new type of FMD vaccine that is more efficient and safe.

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