Analysis of Nucleotide Sequence Encoding VP2 Protein of Infectious Bursal Disease Virus Detected in Korea

국내 분리 닭 전염성 F낭병 바이러스의 VP2 단백질 생산 유전자의 염기서열 분석

  • Kim, Toh-kyung (Gyeongnam Livestock Promotion Institute) ;
  • Yeo, Sang-geon (College of Veterinary Medicine, Gyeongsang National University)
  • Accepted : 2003.07.19
  • Published : 2003.09.25


The VP2 gene of infectious bursal disease virus (IBDV) Chinju which was previously detected in Chinju, Korea was cloned and sequenced to establish the information for the development of genetically engineered vaccines and diagnostic reagents against IBDV. The nucleotide sequence of the entire Chinju VP2 gene consisted of 1,356 bases long encoding 452 amino acids in a single open reading frame (ORF). It consisted of 368 adenine (27.1%), 363 cytosine (26.8%), 339 guanine (25.0%) and 286 thymine (21.1%) residues. The predicted $M_r$ of the Chinju VP2 protein was 48 kDa, and the protein contained 13 phosphorylation sites by protein kinase C, casein kinase II or tyrosine kinase, whereas 3 asparagine-linked glycosylation sites were recognized. The nucleotide sequence of Chinju VP2 ORF had a very close phylogenetic relationship with 98-99% homology to that of the very virulent IBDVs (vvIBDVs) HK46, OKYM, D6948, UK661, UPM97/61 and BD3/99. Also, the Chinju VP2 protein revealed a very close phylogenetic relationship with 99-100% homology to that of these vvIBDVs. The Chinju VP2 protein had 100% amino acid identity in the variable region of residues 206-360 with that of the D6948, HK46, OKYM and UK661, as well as 100% identity in two hypervariable regions of residues 212-224 and 314-324 with those of the D6948, HK46, OKYM, UK661, UPM97/61 and BD3/99. The amino acid sequence of the chinju VP2 protein contained a serine-rich heptapeptide of SWSASGS as in these vvIBDVs.


  1. Akin, A., Wu, C. C. and Lin, T. L. Amplification and cloning of infectious bursal disease virus genomic RNA segments by long and accurate PCR. J. VirolMethods 1999, 82, 55-61
  2. Azad, A, A., Barrett, S. A. and Fahey, K. J. The characterization and molecular cloning of the double-stranded RNA genome of an Australian strain of infectious bursal disease virus. Virol. 1985, 143, 35-44
  3. Bayliss, C. D., Spies, U., Shaw, K., Peters, R W., Papaaeorgiou, A., Muller, H. and Boursnell, M. E. G. A comparison of the sequences of segment A of four infectious bursal disease virus strains and idendfication of a variable region in VP2. J. Gen. Virol. 1990, 71, 1303-1312
  4. Becht, H, Muller. H. and Muller, K K. Comparative studies on structural and antigenic properties of two serotypes of infectious bursal disease virus. J. Gen. Virol. 1988, 69, 631-640
  5. Boot, H. J., ter Huume, A. A. H. M., Hoekman, A.J. W., Peeters, B. P. H. and Gielkens, A. L. J. Rescue of very vinilent and mosaic infectious bursal disease virus from cloned cDNA: VP2 is not the sole detenninant of the very vilrulent phenotype. J. Virol. 2000, 74, 6701-6711
  6. Boot, H. J., ter Huurne, A. A. H. M., Peeters, B. P. H. and Gielkens, A. L. J. Efficient rescue of infectious bursal disease virus from cloned cDNA: evidence for involvement of the 3'-termina1 sequence in genome replication. Virology 1999, 265, 330-341
  7. Box, P. G. High maternal antibodies help chicks beat virulent virus. World Poult. 1989, 53, 17-19
  8. Brown, M. D., Green, P. and Skinner, M, A. VP2 sequences of recent European 'very virulent'isolates of infectious bursal disease virus are closely related to each other but are distinct from those of 'classical' strain. J. Gen. Virol. 1994, 75, 675-680
  9. Brown, M. D. and Skinner, M. A. Coding sequencesof both genome segments of an European 'very virulent' infectious bursal disease virus. Virus Res. 1996, 40, 1-15
  10. Burkhardt, E. and Nuller, H. Susceptibility of chicken blood lymphoblasts and monocytes to infectious bursal disease virus (IBDV). Arch. Virol. 1987, 94, 297-303
  11. Chettle, N. J., Stuart, J. C. and Wyeth, P. J. Outbreak of virulent infectious bursal disease in East Anglia. Vet. Rec. 1989, 125. 271-272
  12. Chong, L. K., Omar, A. R., Yusoff, K., Hair-Bejo, M. and Aini, I. Nucleotidc sequence and phylogenetic analysis of a segrnent of a highly virulent strain ofinfectious bursal disease virus. Acta. Virol. 2001, 45, 217-226
  13. Dobos, P., Hill, B. J., Hallett, R., Kelts, D. T. C., Becht, H. and Teninges, D. Biophysical and biochemical characterization of five animal viruses with bisegmented double-stranded RNA genomes. J. Virol. 1979, 32, 593-605
  14. Domitorio, T. V., Giambrone, J. J. and Duck, L. W. Sequence comparisons of the variable VP2 region of eight infectious bursal disease virus isolates. Avian Dis. 1997, 41, 36-44
  15. Eterradossi, N., PScault, J. P., Drouin, P., Guittet, M, L'Hospitalier, R. and Bennejean, G. Pathogenicity and preliminary antigenic characterization of six infectious bursal disease virus strains isolated in France from acute outbreaks. Zentalbl. Veteinnaermed. 1992, 39, 683-691
  16. Fahey, K. J., Emy, K. M. and Crooks, J. A. Conformational immnunogen on VP2 of infectious bursal disease virus that induces virus-neutralizing antibodies that passively protect chickens. J. Gen.Virol. 1989, 70, 1473-1481
  17. Fahey, K. J., O'DonneIl, I. J. and Azad, A. A.Characterization by Westem blotting of the irnmunogensof infectious bursal disease virus. J. Gen. Virol. 1985,66, 1479-1488
  18. Heine, H. G., Haritou, M., Failla, P., Fahey, K. andAzad, A. A. Sequence analysis and expression of thehost-protective immunogen VP2 of a variant strain ofinfectious bursal disease vims which can circumventvaccination with standard type 1 strains. J. Gen. Virol.1991, 72, 1835-1843
  19. Hudson. P. J., McKem, N. M., Power, R E. andAzad, A. A. Genomic structure of the large RNAsegment of infectious bursal disease virus. NucleicAcids Res. 1986, 14, 5001-5012
  20. Kibenge, F. S. B., Dhillon, A. S. and Russell, R G.Biochemistry and immunology of infectious bursaldisease vims. J. Gen. Virol. 1988, 69, 1757-1775
  21. Kibenee, F. S. B., Jackwood, D. J. and Mercado, C.C. Nucleotide sequence analysis of genoine segment Aof infectious bursal disease virus. J. Gen. Virol. 1990,71, 569-577
  22. Kim, T. K. and Yeo, S. G. Cloning and nucleotideanalysis of segment A gene of infectious bursal disease virus detected in Korea. Virus Genes 2003, 26,97-106
  23. Kwon, H. M., Kim, D. K., Hahn, T. W., Han, J. H.and Jackwood, D. J. Sequence of precursor polyprotein gene (segment A) of infectious bursal disease viruses isolated in Korea. Avian Dis. 2000, 44, 691-696
  24. Lasher, H. N. and Shane, S. M. Infectious bursaldisease. World's Poult. Sci. J. 1994, 50, 133-166
  25. Lim, B. L., Cao, Y., Yu, T. and Mo, C. W. Adaptation of very virulent infectious bursal disease virus to chicken embryonic fibroblasts by site-directed mutagenesis of residues 279 and 284 of viral coat protein VP2. J, Virol. 1999, 73, 2854-2862
  26. Lin, Z., Kato, A., Otaki, V., Nakamura, T., Sasmaz, E. and Ueda, S. Sequence comparisons of a highlyvirulent infectious bursal disease virus prevalent inJapan. Avian Dis. 1993, 37, 315-323
  27. Lukert, P. D. and Saif, Y. M. Infectious BursalDisease. In Diseases of Poultry. lOth ed. Calnek, B.W., Bames, H. J., Beard, C. W., McDougald, L. R.and Saif, Y. M. (eds) pp. 721-738, Iowa StateUniversity Press, Ames, 1997
  28. Mundt, E., Kollner, B. and Kretzschmar, D. VP5 ofinfectious bursal disease virus is not essential for viraliwlication in cell cultun;. J. Viml. 1997, 71, 5647-5651
  29. Murphy, F. A., Gibbs, E. P. J., Horzinek, M. C.and Studdert, M. J. Veterinary Virology, 3rd ed.Academic Press, San Diego, 1999, PP. 405-409
  30. Nagarajan, M. M. and Kibenge, F. S. B. Infectiousbursal disease virus: A review of molecular basis forvariations in antigenicity and virulence. Can. J. Vet.Res. 1997, 61, 81-88
  31. Oppling, V., Muller, a and Becht, J. B Heterogeneityof the antigenic site responsible for the induction ofneutralizing antibodies in infectious bursal diseasevirus. Arch. Virol. 1991, 119, 211-223
  32. Rosenberger, J. K. and Cloud, S. S. Isolation andcharacterization of variant infectious bursal diseaseviruse. In Proceedings of the 123rd Annual Meeting of the Arnerican Veterinary Medical Association, 1986,189, 357
  33. Sambrook, J., Fritsch, E. F. and Maniatis, T.Molecular cloning, In A laboratory mannal. 3rd ed.Cold Spting Harbor Laboratory Press, New York, 2000
  34. Shanna, J. M., Dohms, J. E. and Metz, A. L.Comparative pathogenesis of serotype 1 and variantserotype 1 isolates of infectious bursal disease virusand there effect on humoral and cellular immunecompetence of specific-pathogen-free chickens. AvianDis. 1989, 33, 112-124
  35. Spies, U., Muller, H. and Becht, H. Nucleotidesequence of infectious bursal disease virus genomesegment A delineates two major open reading frames.Nucleic Acids Res. 1989, 17, 7982
  36. Spies, U., Muller, H. and Becht, H. Properties ofRNA polyrnerase activity associated with infectiousbursal disease virus and characterization of its reactionproducts. Virus Res. 1987, 8, 127-140
  37. Tacken, M. G. J., Rottier, P. J. M., Gielkens, A. L.J. and Peeters, B. P. H. Interactions in vivo betweenthe proteins of infectious bursal disease virus: capsidprotein VP3 interacts with the RNA-dependent RNApoltmerase, VPl. J. Gen. Virol. 2000, 81, 209-218
  38. Tsai, H. J. and Lu, Y. S. Epizoodology of infectiousbursal disease in Taiwan in 1992. J. Chin. Soc. Vet.Sci. 1993, 19, 249-258
  39. Vakharia, V. N., He, J., Ahamed, B. and Snyder, D.B. Molecular basis of antigenic variation in infectiousbursa disease virus. Virus Res. 1994, 31, 265-273
  40. Van Den Berg, T. P., Gonze, M. and Meulemans,G. Acute infectious bmsal disease in poultry: isolationand characterization of highly virulent strain. AvianPathol. 1991, 20, 133-143
  41. Yamaguchi, T., Ogawa, M., Inoshinia, Y., Miyoshi,M., Fukushi, H. and Hirai, K. Identification ofsequence changes responsible for the attenuation ofhighly virulent infectious bursal disease virus. Virology1996, 223, 219-223