Bovine viral diarrhea virus E0 protein amino acid and preparation method thereof

Abstract

The invention provides a bovine viral diarrhea virus E0 protein amino acid and a preparation method thereof. The amino acid sequence of the bovine viral diarrhea virus E0 protein amino acid is represented by SEQ ID NO:3. The preparation method comprises the following steps:S1, obtaining a BVDV NADL whole genome sequence from NCBI GenBank, and finding out a BVDV E0 gene sequence from the BVDV NADLwhole genome sequence; S2, optimizing and synthesizing the E0 gene sequence obtained in step S1; and S3, selecting a prokaryotic expression vector pMal-c5X with an MBP label, connecting the E0 gene with the pMal-c5X, inserting an E0 fragment into the pMal-c5X, converting the connected expression vector pMAL-c5x-E0 into an expression host bacterium BL21 (DE3), inducing the expression of the obtained recombinant protein, and separating and purifying the target protein. The target gene E0 is optimized, and is subjected to fusion expression with the MBP tag protein to finally achieve the efficientsoluble expression of the E0 protein, so the bovine viral diarrhea virus E0 protein amino acid has the advantages of high expression yield, good solubility, and convenience in expression protein purification; and the solubility of the Escherichia coli expression protein can be improved during the fusion expression of the foreign protein and the MBP so as to provide convenience for the purification of the expression protein.

Description

technical field [0001] The invention relates to a bovine viral diarrhea virus E0 protein amino acid and a preparation method. Background technique [0002] Bovine viral diarrhea virus (BVDV) is the causative agent of bovine viral diarrhea (BVD) and a member of the Pestivirus genus in the family Flaviviridae; it can infect cattle, sheep, pigs, and camels Animals such as deer, alpacas and wallabies threaten the cattle, dairy and meat industries around the world, with the emergence of more and more highly virulent virus strains causing increasing economic losses over the past few decades . BVD is a pantropic viral infection that affects reproductive and growth performance and causes immunosuppression in cattle. Reproductive disorders caused by BVDV mainly include miscarriage, death of early fertilized eggs and fatal defects. Performance effects include reduced feed conversion, lower body carcass mass, and reduced milk quality. The main clinical manifestations are diarrhea, ...

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Problems solved by technology

[0004] At present, some BVDV E0 protein expression in vitro has been reported, but most of the expressed E0 protein is in the form of inclusion body (literature). Although the expression level of the inclusion body form is high, this protein form has no activity, and the late recovery process is complicated and tortuous, and the purification is difficult Difficult; another part reports that the soluble E0 protein can be expressed using the insect baculovirus expression system, but the expression level is not high, and the operation is not as convenient as the prokaryotic expression process, and the cost is also high, which is not conducive to the later mass production and use

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