Applied Microbiology and Biotechnology

, Volume 68, Issue 2, pp 220–227

Expression of SARS-coronavirus nucleocapsid protein in Escherichia coli and Lactococcus lactis for serodiagnosis and mucosal vaccination

  • Huadong Pei
  • Jingfang Liu
  • Yun Cheng
  • Chaomin Sun
  • Chen Wang
  • Yueping Lu
  • Jie Ding
  • Jian Zhou
  • Hua Xiang
Applied Genetics and Molecular Biotechnology

DOI: 10.1007/s00253-004-1869-y

Cite this article as:
Pei, H., Liu, J., Cheng, Y. et al. Appl Microbiol Biotechnol (2005) 68: 220. doi:10.1007/s00253-004-1869-y

Abstract

The nucleocapsid (N) protein of the severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) is an important antigen for the early diagnosis of SARS and the development of vaccines. It was expressed in Escherichia coli as a fusion with human glutathione S-transferase (hGST) and was confirmed by Western blotting analysis. This recombinant N protein (hGST-N) was purified and used to measure the SARS-CoV N-specific antibody in the sera of eight SARS patients by enzyme-linked immunosorbent assay. Specific antibody response to this purified recombinant N protein was 100% positive in the SARS patients’ sera, while none of the control sera from 30 healthy people gave a positive reaction in the same assay. The SARS-CoV N protein was also expressed in Lactococcus lactis in the cytoplasm or secreted into the medium. The N-producing strain MG1363/pSECN and the purified hGST-N protein were respectively administered to mice, either orally or intranasally. Results indicated that orally delivered MG1363/pSECN induced significant N-specific IgG in the sera. In conclusion, our work provides a novel strategy to produce the SARS-CoV N protein for serodiagnosis and for L. lactis-based mucosal vaccines.

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Huadong Pei
    • 1
    • 2
  • Jingfang Liu
    • 1
    • 2
  • Yun Cheng
    • 1
  • Chaomin Sun
    • 1
    • 2
  • Chen Wang
    • 3
  • Yueping Lu
    • 3
  • Jie Ding
    • 3
  • Jian Zhou
    • 1
  • Hua Xiang
    • 1
  1. 1.State Key Laboratory of Microbial Resources, Institute of MicrobiologyChinese Academy of SciencesBeijingP.R. China
  2. 2.Graduate school of the Chinese Academy of SciencesBeijingP.R. China
  3. 3.Chao-Yang Hospital Affiliated to Capital University of Medical SciencesBeijingP.R. China