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- Title
Expression and immunogenicity of non-structural protein 8 of porcine epidemic diarrhea virus.
- Authors
Hong Chen; Jiawu Wan; Meihua Wei; Ping Liu; Lingbao Kong; Xiu Xin
- Abstract
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 pMALc2x-maltose binding protein (MBP) and pET-28a (+). Subsequently, the optimization of expression conditions in Escherichia coli, including induced temperature, time and isopropyl ß-D-thiogalactopyranoside concentration were performed to obtain a stable expression of MBPnsp8 and nsp8. The nsp8 fused with MBP increased the water solubility of the expressed products. Target proteins were further purified from E. coli culture and their immunogenicities were evaluated in vivo 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.
- Subjects
ESCHERICHIA coli; IMMUNOGENETICS; PORCINE epidemic diarrhea virus; MICROBIAL virulence; MALTOSE binding proteins
- Publication
Veterinary Research Forum, 2024, Vol 15, Issue 2, p65
- ISSN
2008-8140
- Publication type
Article
- DOI
10.30466/vrf.2023.2009322.3977