Expression and immunogenicity of non-structural protein 8 of porcine epidemic diarrhea virus.

Mar 11, 2024

Experts: Hong Chen, Jiawu Wan, Meihua Wei, Ping Liu, Lingbao Kong, Xiu Xin

The non-structural protein (nsp) 8 of the porcine epidemic diarrhea virus (PEDV) is highly stable across different PEDV strains and plays an important role in PEDV virulence. In current study, nsp8 prokaryotic expression vectors were constructed based on parental vectors pMAL-c2x-maltose binding protein (MBP) and pET-28a (+). Subsequently, the optimization of expression conditions in , including induced temperature, time and isopropyl β-D-thiogalactopyranoside concentration were performed to obtain a stable expression of MBP-nsp8 and nsp8. The nsp8 fused with MBP increased the water solubility of the expressed products. Target proteins were further purified from culture and their immunogenicities were evaluated by mice. The antibody titers of serum from nsp8 immunized mice were up to 1:7,750,000 when measured by indirect enzyme-linked immunosorbent assay; meanwhile, the mice immunized with MBP-nsp8 gave an antibody titer reaching 1:1,000,000. In all, the expression and purification system of PEDV nsp8 and MBP-nsp8 were successfully established in this work and a strong immune response was elicited in mice by both purified nsp8 and MBP-nsp8, providing a basis for the study of the structure and function of PEDV nsp8.
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ABOUT THE EXPERTS

Hong Chen, Jiawu Wan, Meihua Wei, Ping Liu, Lingbao Kong, Xiu Xin

Hong Chen

Institute of Pathogenic Microbiology, College of Biological Science and Engineering, Jiangxi Agricultural University, Nanchang, China.

Nanchang Key Laboratory of Animal Virus and Genetic Engineering, College of Biological Science and Engineering, Jiangxi Agricultural University, Nanchang, China.

Jiawu Wan

Institute of Pathogenic Microbiology, College of Biological Science and Engineering, Jiangxi Agricultural University, Nanchang, China.

Nanchang Key Laboratory of Animal Virus and Genetic Engineering, College of Biological Science and Engineering, Jiangxi Agricultural University, Nanchang, China.

Meihua Wei

Institute of Pathogenic Microbiology, College of Biological Science and Engineering, Jiangxi Agricultural University, Nanchang, China.

Nanchang Key Laboratory of Animal Virus and Genetic Engineering, College of Biological Science and Engineering, Jiangxi Agricultural University, Nanchang, China.

Ping Liu

Institute of Pathogenic Microbiology, College of Biological Science and Engineering, Jiangxi Agricultural University, Nanchang, China.

Nanchang Key Laboratory of Animal Virus and Genetic Engineering, College of Biological Science and Engineering, Jiangxi Agricultural University, Nanchang, China.

Lingbao Kong

Institute of Pathogenic Microbiology, College of Biological Science and Engineering, Jiangxi Agricultural University, Nanchang, China.

Nanchang Key Laboratory of Animal Virus and Genetic Engineering, College of Biological Science and Engineering, Jiangxi Agricultural University, Nanchang, China.

Xiu Xin

Institute of Pathogenic Microbiology, College of Biological Science and Engineering, Jiangxi Agricultural University, Nanchang, China.

Nanchang Key Laboratory of Animal Virus and Genetic Engineering, College of Biological Science and Engineering, Jiangxi Agricultural University, Nanchang, China.