Prokaryotic Expression of Gene VP2 of Porcine Parvovirus Type 1 and the Reactinogenicity Analysis of the Expressed Protein

doi:10.13560/j.cnki.biotech.bull.1985.2017.03.024

Abstract

Abstract: This experiment is aimed to study the antigenicity of VP2 protein（amino acid 155-439）of porcine parvovirus type 1（PPV1）for laying a base for the development of detecting PPV1. The 849 bp target fragment was amplified using the DNA of strain AV31of PPV1 as the template. The product was cloned into pET30a（+）vector,and recombinant plasmid pET30a-PPV1-VP2（amino acid 155-439）was constructed,then transferred into Escherichia coli BL21（DE3）for the 6 h induced expression by IPTG. The recombinant protein was purified by Ni-NTA,and refolded by different concentration of urea. The results of SDS-PAGE showed that the gene VP2 was successfully expressed in E. coli BL21（DE3）with a relative molecular weight of 39 kD. The results of Western blot showed that this recombined protein specifically reacted with PPV1 positive serum,while no cross reaction with NA-PRRSV and PCV2 positive serum. This study achieved the aims：the recombinant vector pET30a-PPV1-VP2 was successfully constructed,and the gene was successfully expressed in E. coli,and the purified and re-folded protein demonstrated promising reactinogenicity .

Key words: porcine parvovirus type 1（PPV1）, VP2 gene, prokaryotic expression, reactinogenicity

OU Yun-wen, MA Xiao-yuan, ZHANG Jie, DING Yao-zhong, ZHANG Yong-guang, JIA Ning. Prokaryotic Expression of Gene VP2 of Porcine Parvovirus Type 1 and the Reactinogenicity Analysis of the Expressed Protein[J]. Biotechnology Bulletin, 2017, 33(3): 169-174.

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