Chemically-synthesized porcine circo-virus (PCV) protein gene, modification method and applications thereof

Abstract

The invention discloses a chemically-synthesized porcine circo-virus (PCV) protein gene, a modification method, and applications thereof. The preparation of the gene comprises the following steps: obtaining and modifying a PCV2ORF2 gene sequence, designing and synthesizing a primer, synthesizing PCV2ORF2 short-section DNA, synthesizing the full-length gene, determining the sequence of the full-length gene, constructing an expression vector, and expressing a target gene in pichia pastoris. The method takes the PTDS method and PAS method as the references, and reduces the gene synthesis cost, moreover, the procedure of the method is simple, and only comprises two steps of PCR, the error rate is low, and the synthesis period is short. During the whole chemical synthesis process, high fidelity Taq enzyme is used, and thus the mistakenly matched base sequences can be corrected during the synthesis process so as to maximally reduce the generation of mistakenly matched base pair. The gene is cloned to a yeast expression vector to construct a recombinant expression plasmid, and the expression product extracted from large-scale fermentation of the recombinant expression plasmid can be used as the diagnosis antigen for PCV2 antibody detection, and can also be used to prepare subunit vaccine or nucleic acid vaccine.

Description

technical field [0001] The invention relates to a protein, in particular to a chemically synthesized porcine circovirus Cap protein gene and its transformation method and application. Background technique [0002] Porcine circovirus (Porcine circovirus, PCV) is divided into two genotypes, PCV1 and PCV2, of which PCV2 is pathogenic and can cause postweaning multisystemic wasting syndrome (postweaning multisystemic wasting syndrome, PMWS), or fattening pigs The main cause of porcine dermatitis and nephrotic syndrome (Porcine Dermatitis and Nephropathy Syndrome, PDNS). Because the disease is an immunosuppressive disease, it can cause severe secondary infection and endanger the health of pig herds. It is another major disease that seriously endangers the global pig industry after porcine reproductive and respiratory syndrome. PCV2 in large-scale pig herds in my country The infection rate exceeds 90%. In the past two years, there are 4 porcine circovirus vaccines that have been ...

AI Technical Summary

Problems solved by technology

The difficulty in the research of porcine circovirus type 2 genetically engineered vaccines is how to improve the protein yield and immunogenicity. There have been many studies in the world using the products expressed by E. coli, baculovir

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