Identification of a conserved B-cell epitope in the equine arteritis virus (EAV) N protein using the pepscan technique

Virus Genes - Tập 47 - Trang 292-297 - 2013
Shihua Zhao1, Ting Qi1, Wei Guo1, Gang Lu1, Wenhua Xiang1
1Division of Livestock Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, People’s Republic of China

Tóm tắt

The nucleocapsid (N) gene of equine arteritis virus (EAV) is highly conserved between isolates, and the N protein is an important antigen that induces immunity when horses are infected with EAV. This study describes the identification of a linear B-cell epitope on the N protein using the pepscan technique with a monoclonal antibody (mAb) 2B1 directed against the N protein. The N protein was divided into 11 overlapping peptides, each containing 16 amino acids associated with six overlapping amino acids. The fragments were expressed as MBP fusion proteins that were then used to probe the 2B1 mAb. The minimal epitope sequence was confirmed step-by-step using single amino acid residue deletion. One completely conserved linear epitope (38KPPAQP43) was identified that matched with EAV-positive serum in Western blots, thereby revealing the importance of these six amino acids of the epitope for antibody-epitope binding activity. This finding not only contributes to our understanding of the antigenic structure of the N protein of EAV but also has potential for the development of diagnostic techniques.

Tài liệu tham khảo

U.B. Balasuriya, N.J. MacLachlan, Vet. Immunol. Immunopathol 102, 107–129 (2004) J.T. Bryans, M.E. Crowe, E.R. Doll, W.H. McCollum, Cornell Vet 47, 3–41 (1957) J.T. Bryans, E.R. Doll, R.E. Knappenberger, Cornell Vet 47, 69–75 (1957) A.L. Glaser, A.A. de Vries, E.J. Dubovi, J. Gen. Virol 76(Pt 9), 2223–2233 (1995) Y. Fukunaga, W.H. McCollum, Am. J. Vet. Res 38, 2043–2046 (1977) Y.Y. Go, E.J. Snijder, P.J. Timoney, U.B. Balasuriya, Clin. Vaccine Immunol 18, 268–279 (2011) E. Weiland, S. Bolz, F. Weiland, W. Herbst, M.J. Raamsman, P.J. Rottier, A.A. De Vries, J. Clin. Microbiol 38, 2065–2075 (2000) F. Miszczak, L. Legrand, U.B. Balasuriya, B. Ferry-Abitbol, J. Zhang, A. Hans, G. Fortier, S. Pronost, A. Vabret, Virology 423, 165–174 (2012) E.D. Chirnside, P.M. Francis, J.A. Mumford, Virus Res 39, 277–288 (1995) A. Kheyar, S. Martin, G. St-Laurent, P.J. Timoney, W.H. McCollum, D. Archambault, Clin. Diagn. Lab. Immunol 4, 648–652 (1997) A.A. de Vries, E.D. Chirnside, M.C. Horzinek, P.J. Rottier, J. Virol 66, 6294–6303 (1992) M.N. Dharmasena, S.J. Krebs, R.K. Taylor, Microbiology 155, 2353–2364 (2009) M.J. Gubbins, J.D. Berry, C.R. Corbett, J. Mogridge, X.Y. Yuan, L. Schmidt, B. Nicolas, A. Kabani, R.S. Tsang, FEMS Immunol. Med. Microbiol 47, 436–443 (2006) R. Wieringa, A.A. de Vries, J. van der Meulen, G.J. Godeke, J.J. Onderwater, H. van Tol, H.K. Koerten, A.M. Mommaas, E.J. Snijder, P.J. Rottier, J. Virol 78, 13019–13027 (2004) J. Zhang, Y.Y. Go, C.M. Huang, B.J. Meade, Z. Lu, E.J. Snijder, P.J. Timoney, U.B. Balasuriya, Clin. Vaccine Immunol 19, 1312–1321 (2012) E. Starik, A. Ginter, P. Coppe, J. Vet. Med. B Infect. Dis. Vet. Public Health 48, 1–9 (2001) Y.J. Zhou, H. Yu, Z.J. Tian, J.X. Liu, T.Q. An, J.M. Peng, G.X. Li, Y.F. Jiang, X.H. Cai, Q. Xue, M. Wang, Y.F. Wang, G.Z. Tong, Vet. Microbiol 138, 1–10 (2009) S. Mankoc, P. Hostnik, J. Grom, I. Toplak, I. Klobucar, M. Kosec, D. Barlic-Maganja, J. Virol. Methods 146, 341–354 (2007)