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.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
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
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
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
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
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
S.K. Wootton E.A. Nelson D. Yoo (1998) Clin Diagn Lab Immunol 5 773–779 Occurrence Handle1:CAS:528:DyaK1cXnvVCjtrc%3D Occurrence Handle9801333
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
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
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
G.P. Smith (1985) Science 228 1315–1317
D.R. Burton (1995) Immunotechnology 1 87–94 Occurrence Handle10.1016/1380-2933(95)00013-5 Occurrence Handle1:CAS:528:DyaK2MXotVOnurs%3D Occurrence Handle9373337
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
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
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
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
V.F. la de Cruz A.A. Lal T.F. McCutchan (1988) J Biol Chem 263 4318–4322 Occurrence Handle2450091
S.F. Parmley G.P. Smith (1989) Adv Exp Med Biol 251 215–218 Occurrence Handle1:CAS:528:DyaK3cXktVChs74%3D Occurrence Handle2481962
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
Y. Wan G.P. Dai Y. Wang (1999) J Third Military Med Uni 21 837–839 Occurrence Handle1:CAS:528:DC%2BD3cXjt1Cjsw%3D%3D
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
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
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
M.B. Oleksiewicz A. Bøtner P. Normann (2002) J Gen Virol 83 1407–1418 Occurrence Handle1:CAS:528:DC%2BD38XktlGitLw%3D Occurrence Handle12029156
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
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
J. Sambrook E.F. Fritsch T. Maniatis (1989) Molecular Cloning: A Laboratory Manual EditionNumber2 Cold Spring Harbor Laboratory Cold Spring Harbor, NY
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
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
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
Author information
Authors and Affiliations
Corresponding author
Rights 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
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11262-005-2203-1