Skip to main content
SpringerLink
Log in

Identification of a Novel B Cell Epitope on the Nucleocapsid Protein of Porcine Reproductive and Respiratory Syndrome Virus by Phage Display

  • Published:
Virus Genes Aims and scope Submit manuscript

Abstract

A phage display peptide library targeting the nucleocapsid (N) protein of porcine reproductive and respiratory syndrome virus (PRRSV) strain CH-1a was generated and used for epitope mapping. After 3 rounds of biopanning with the monoclonal antibody (MAb) N3H2 directed against the N protein, 3 positive phages were screened and sequenced. These phages share a consensus sequence, IQTAFNQGA, which corresponds to the amino acid (AA) 79–87 segment of the CH-1a N protein. A small DNA fragment coding for IQTAFNQGA was expressed as a fusion product, and reacted to N3H2 in Western blots and indirect ELISA. Four truncated peptides (IQTAFNQG, IQTAFNQ, QTAFNQGA, and TAFNQGA) expressed as GST fusion products failed to react with N3H2. The sequences around the N3H2-binding site among the N proteins of 57 PRRSV strains were compared. Our results indicate that the IQTAFNQGA motif is highly conserved among North American and European isolates. We concluded that the precisely defined nona-peptide epitope is a novel conserved Linear B cell epitope on the N protein of PRRSV.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. D.A. Benfield E. Nelson J.E. Collins L. Harris S.M. Goyal D. Robinson W.T. Christianson R.B. Morrison D.E. Gorcyca D.W. Chladek (1992) J Vet Diagn Invest 4 127–133 Occurrence Handle1:STN:280:By2A3MvgvFE%3D Occurrence Handle1616976

    CAS  PubMed  Google Scholar 

  2. G. Wensvoort C. Tepstra J.M.A. Pol E.A. ter Laak M. Bloemraad E.P. de Kluyver C. Kragten L. van Buiten A. den Besten F. Wagenaar J.M. Broekhuijsen P.L.J.M. Moonen T. Zestra E.A. de Boer H.J. Tibben M.F. de Jong P. van Veld G.J.R. Groenland J.A. van Gennep M.T. Voets J.H.M. Verheijden Braamskamp (1991) J., Vet Q 13 121–130

    Google Scholar 

  3. J.E. Collins D.A. Benfield W.T. Christianson J.E. Harris L. Hennings J.C. Shaw D.P. Goyal S.M. McCullough S. Morrison R.B. Joo H.S. Gorcyca D.W. Chladek (1992) J Vet Diagn Invest 4 117–126 Occurrence Handle1:STN:280:By2A3MvgvFA%3D Occurrence Handle1616975

    CAS  PubMed  Google Scholar 

  4. E.A. Nelson J.K. Christopher-Hennings T. Drew G. Wensvoort J. Collins D.A. Benfield (1993) J Clin Microbiol 31 3184–3189 Occurrence Handle1:STN:280:ByuC2c%2FpvV0%3D Occurrence Handle7508455

    CAS  PubMed  Google Scholar 

  5. G. Wensvoort E.P. de Kluyver E.A. Luitjze A. den Besten L. Harris J.E. Collins W.T. Christianson D. Chladek (1992) J Vet Diagn Invest 4 134–138 Occurrence Handle1:STN:280:By2A3MvgvFI%3D Occurrence Handle1616977

    CAS  PubMed  Google Scholar 

  6. S.K. Wootton E.A. Nelson D. Yoo (1998) Clin Diagn Lab Immunol 5 773–779 Occurrence Handle1:CAS:528:DyaK1cXnvVCjtrc%3D Occurrence Handle9801333

    CAS  PubMed  Google Scholar 

  7. J.J. Meulenberg A.P. van Nieuwstadt A. van Essen-Zandbergen J.N. Bos-de Ruijter J.P. Langeveld R.H. Meloen (1998) Virology 252 106–114 Occurrence Handle10.1006/viro.1998.9436 Occurrence Handle1:CAS:528:DyaK1cXotFSitbw%3D Occurrence Handle9875321

    Article  CAS  PubMed  Google Scholar 

  8. L. Yang K.J. Yoon Y. Li J.H. Lee J.J. Zimmerman M.L. Frey K.M. Harmon K.B. Platt (1999) Arch Virol 144 525–546 Occurrence Handle10.1007/s007050050523 Occurrence Handle1:CAS:528:DyaK1MXjs12rt7k%3D Occurrence Handle10226618

    Article  CAS  PubMed  Google Scholar 

  9. S. Wootton G. Koljesar L. Yang K.J. Yoon D. Yoo (2001) Clin Diagn Lab Immunol 8 598–603 Occurrence Handle10.1128/CDLI.8.3.598-603.2001 Occurrence Handle1:CAS:528:DC%2BD3MXktVOms7Y%3D Occurrence Handle11329465

    Article  CAS  PubMed  Google Scholar 

  10. G.P. Smith (1985) Science 228 1315–1317

    Google Scholar 

  11. D.R. Burton (1995) Immunotechnology 1 87–94 Occurrence Handle10.1016/1380-2933(95)00013-5 Occurrence Handle1:CAS:528:DyaK2MXotVOnurs%3D Occurrence Handle9373337

    Article  CAS  PubMed  Google Scholar 

  12. F. Bugli N. Mancini C.Y. Kang C.D. Campli A. Grieco A. Manzin A. Gabrielli A. Gasbarrini G. Fadda P.E. Varaldo M. Clementi R. Burioni (2001) J Virol 75 9986–9990 Occurrence Handle10.1128/JVI.75.20.9986-9990.2001 Occurrence Handle1:CAS:528:DC%2BD3MXnt1Ggtbo%3D Occurrence Handle11559832

    Article  CAS  PubMed  Google Scholar 

  13. L.F. Wang D.H. DuPlessis J.R. White A.D. Hyatt B.T. Eaton (1995) J Immunol Method 178 1–12 Occurrence Handle10.1016/0022-1759(94)00235-O Occurrence Handle1:CAS:528:DyaK2MXjt1WjsL0%3D

    Article  CAS  Google Scholar 

  14. J. Robben K. Hertveldt E. Bosmans G. Volckaert (2002) J Biol Chem 277 17544–17547 Occurrence Handle10.1074/jbc.M110275200 Occurrence Handle1:CAS:528:DC%2BD38XktVCgtb8%3D Occurrence Handle11825896

    Article  CAS  PubMed  Google Scholar 

  15. A.M. Coley N.V. Campanale J.L. Casey A.N. Hodder P.E. Crewther R.F. Anders L.M. Tilley M. Foley (2001) Protein Eng 14 691–698 Occurrence Handle10.1093/protein/14.9.691 Occurrence Handle1:CAS:528:DC%2BD3MXovVOgsbc%3D Occurrence Handle11707616

    Article  CAS  PubMed  Google Scholar 

  16. V.F. la de Cruz A.A. Lal T.F. McCutchan (1988) J Biol Chem 263 4318–4322 Occurrence Handle2450091

    PubMed  Google Scholar 

  17. S.F. Parmley G.P. Smith (1989) Adv Exp Med Biol 251 215–218 Occurrence Handle1:CAS:528:DyaK3cXktVChs74%3D Occurrence Handle2481962

    CAS  PubMed  Google Scholar 

  18. B.Q. Guo Z.S. Chen W.X. Liu Y.Z. Cui L.D. Kong (1996) Chin J Infect Dis Anim Poult 18 1–5

    Google Scholar 

  19. Y. Wan G.P. Dai Y. Wang (1999) J Third Military Med Uni 21 837–839 Occurrence Handle1:CAS:528:DC%2BD3cXjt1Cjsw%3D%3D

    CAS  Google Scholar 

  20. M.B. Oleksiewicz A. Bøtner K.G. Madsen T. Storgaard (1998) Vet Microbiol 64 7–22 Occurrence Handle10.1016/S0378-1135(98)00254-5 Occurrence Handle1:CAS:528:DyaK1cXns1Oisrg%3D Occurrence Handle9874099

    Article  CAS  PubMed  Google Scholar 

  21. H.S. Nielsen T. Storgaard M.B. Oleksiewicz (2000) Vet Microbiol 76 221–228 Occurrence Handle10.1016/S0378-1135(00)00258-3 Occurrence Handle1:CAS:528:DC%2BD3cXmtVCls7c%3D Occurrence Handle10973697

    Article  CAS  PubMed  Google Scholar 

  22. M.B. Oleksiewicz A. Bøtner P. Toft P. Normann T. Storgaard (2001) J Virol 75 3277–3290 Occurrence Handle10.1128/JVI.75.7.3277-3290.2001 Occurrence Handle1:CAS:528:DC%2BD3MXitVGiur8%3D Occurrence Handle11238854

    Article  CAS  PubMed  Google Scholar 

  23. M.B. Oleksiewicz A. Bøtner P. Normann (2002) J Gen Virol 83 1407–1418 Occurrence Handle1:CAS:528:DC%2BD38XktlGitLw%3D Occurrence Handle12029156

    CAS  PubMed  Google Scholar 

  24. Y. Wan Y. Wu J. Bian X.Z. Wang W. Zhou Z.C. Jia Y. Tan L. Zhou (2001) Vaccine 9 2918–2923 Occurrence Handle10.1016/S0264-410X(00)00561-2

    Article  Google Scholar 

  25. S.P. Zhang G. Zubay E. Goldman (1991) Gene 105 61–72 Occurrence Handle10.1016/0378-1119(91)90514-C Occurrence Handle1:CAS:528:DyaK38XjsFCjtw%3D%3D Occurrence Handle1937008

    Article  CAS  PubMed  Google Scholar 

  26. J. Sambrook E.F. Fritsch T. Maniatis (1989) Molecular Cloning: A Laboratory Manual EditionNumber2 Cold Spring Harbor Laboratory Cold Spring Harbor, NY

    Google Scholar 

  27. S.K. Wootton D. Yoo (2003) J Virol 77 4546–4557 Occurrence Handle10.1128/JVI.77.8.4546-4557.2003 Occurrence Handle1:CAS:528:DC%2BD3sXivFWis7o%3D Occurrence Handle12663761

    Article  CAS  PubMed  Google Scholar 

  28. D.N. Doan T. Dokland (2003) Structure (Camb) 11 1445–1451 Occurrence Handle10.1016/j.str.2003.09.018 Occurrence Handle1:CAS:528:DC%2BD3sXovVels7g%3D

    Article  CAS  Google Scholar 

  29. J.I. Casal M.J. Rodriguez J. Sarraseca J. Garcia J. Plana-Duran A. Sanz (1998) Adv Exp Med Biol 440 469–477 Occurrence Handle1:CAS:528:DyaK1cXmvFKntrw%3D Occurrence Handle9782317

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, CAAS, 427 Maduan Street, 150001, Harbin, People’s Republic of China

    Tong-Qing An, Yan-Jun Zhou, Hua-Ji Qiu, Guang-Zhi Tong, Yun-Feng Wang, Jin-Xia Liu & Jin-Yu Yang

Authors
  1. Tong-Qing An
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yan-Jun Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hua-Ji Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Guang-Zhi Tong
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yun-Feng Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jin-Xia Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jin-Yu Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guang-Zhi Tong.

Rights and permissions

Reprints and permissions

About this article

Cite this article

An, TQ., Zhou, YJ., Qiu, HJ. et al. Identification of a Novel B Cell Epitope on the Nucleocapsid Protein of Porcine Reproductive and Respiratory Syndrome Virus by Phage Display. Virus Genes 31, 81–87 (2005). https://doi.org/10.1007/s11262-005-2203-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11262-005-2203-1

Keywords

Search

Navigation