Virus Genes

, Volume 43, Issue 3, pp 335–341 | Cite as

Bacterial expression of antigenic sites A and D in the spike protein of transmissible gastroenteritis virus and evaluation of their inhibitory effects on viral infection

  • Fandan Meng
  • Zeping Zhao
  • Guangxing Li
  • Siqingaowa Suo
  • Na Shi
  • Jiechao Yin
  • Dante Zarlenga
  • Xiaofeng Ren
Article
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Abstract

The spike (S) protein is a key structural protein of coronaviruses including, the porcine transmissible gastroenteritis virus (TGEV). The S protein is a type I membrane glycoprotein located in the viral envelope and is responsible for mediating the binding of viral particles to specific cell receptors and therefore specific cell types. It is also an important immune target for the host in neutralizing the virus. Four antigenic sites A, B, C, and D that reside near the N-terminal domain have been defined in the S protein. Of these, the region encoding antigenic sites A and to a lesser extent D, herein defined as S-AD, are most critical in eliciting host neutralizing antibodies. Herein, we enzymatically amplified, cloned, and expressed the S-AD fragment from TGEV in the prokaryotic expression vector, pET-30a. Maximum protein expression was achieved at 30°C over a 5-h period post-induction. Rabbit polyclonal antiserum was generated using recombinant S-AD (rS-AD) protein. In contrast to prior studies showing no activity with bacterially produced S protein, results indicated that polyclonal serum recognized TGEV-infected cells and reduced infection by 100%. Furthermore, the truncated rS-AD peptide was able to bind to the surface of cells from swine testes in a competitive manner and completely inhibit viral infection.

Keywords

Coronavirus TGEV S protein Antibody Cell infection 
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Notes

Acknowledgments

We thank Dr. Luis Enjuanes, CSIC, Madrid, Spain, for providing TGEV and Drs. Georg Herrler and Christel Schwegmann-Wessels, University of Veterinary Medicine Hannover, Germany, for providing the S gene. This study is funded in part by the National Natural Science Foundation of China (30972195). Additional funding is supported by the Open Project Program of Beijing Key Laboratory of Traditional Chinese Veterinary Medicine at Beijing University of Agriculture (TCVM-201103); the Program for New Century Excellent Talents at the Heilongjiang Provincial University (1155–NCET–005) and supported by the New Century Excellent Talents program from the Ministry of Education of P. R. China.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Fandan Meng
    • 1
  • Zeping Zhao
    • 2
  • Guangxing Li
    • 1
  • Siqingaowa Suo
    • 1
  • Na Shi
    • 1
  • Jiechao Yin
    • 1
  • Dante Zarlenga
    • 3
  • Xiaofeng Ren
    • 1
  1. 1.College of Veterinary MedicineNortheast Agricultural UniversityHarbinChina
  2. 2.College of Animal Science and TechnologyChina Agricultural UniversityBeijingChina
  3. 3.Animal Parasitic Diseases LaboratoryAgricultural Research Service, U.S. Department of AgricultureBeltsvilleUSA

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