Prokaryotic expression and bioinformatics analysis of PPV7 Cap gene

doi:10.3969/j.issn.1004-1524.2020.02.03

›› 2020, Vol. 32 ›› Issue (2): 200-209.DOI: 10.3969/j.issn.1004-1524.2020.02.03

• Animal Science • Previous Articles Next Articles

Prokaryotic expression and bioinformatics analysis of PPV7 Cap gene

WANG Xiaopeng¹, ZHAO Liang¹, LIU Zimin¹, BAI Caixia¹, YANG Kankan¹, ZHANG Da¹, SUN Pei¹, JIANG Shudong¹, LI Yongdong^2,*, WANG Yong^1,*

1.College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036,China;
2.Ningbo Municipal Center for Disease Control and Prevention, Ningbo 315010,China

Received:2019-07-01 Online:2020-02-25 Published:2020-03-13

Abstract

Abstract: The porcine parvovirus 7 (PPV7) Cap gene was amplified by PCR,and the amplified product was cloned into the prokaryotic expression vector pGEX-6P-1.The recombinant plasmid pGEX-6P-1-Cap was identified by sequencing and the recombinant plasmid was transformed into E. coli Rosetta cells and induced by IPTG. The expression products were then identified by SDS-PAGE and Western-blot. At the same time, physical and chemical properties,secondary structure,signal peptide and transmembrane domain of the gene encoded protein were predicted using the bioinformatics softwares. The results showed that the recombinant plasmid pGEX-6P-1-Cap was successfully constructed and the expressed Cap protein was about 78 ku, which was consistent with the expectation. The induced Cap protein could react specifically with GST monoclonal antibody and had good reactivity. Bioinformatics analysis results showed that the Cap protein was stable hydrophilic protein,with 62.90% irregular curl(c)structure in the secondary structure, 11.09%,4.05% and 21.96% of α-helix(h), β-turn(t)and extended(e)chain respectively. Moreover, there was no signal peptide area and transmembrane domain. There were 62 phosphorylation sites,12 B cell antigen epitopes,2 CTL epitopes and 3 Th epitopes. In this study, PPV7 Cap protein was successfully expressed and its biological characteristics were predicted, which could provide a reference for the study of biological function of PPV7 Cap protein.

Key words: porcine parvovirus 7, Cap protein, prokaryotic expression, bioinformatics

CLC Number:

S855.3

WANG Xiaopeng, ZHAO Liang, LIU Zimin, BAI Caixia, YANG Kankan, ZHANG Da, SUN Pei, JIANG Shudong, LI Yongdong, WANG Yong. Prokaryotic expression and bioinformatics analysis of PPV7 Cap gene[J]. , 2020, 32(2): 200-209.

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Prokaryotic expression and bioinformatics analysis of PPV7 Cap gene

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