1887

Abstract

Porcine circovirus 2 (PCV2), a single-stranded DNA virus associated with post-weaning multisystemic wasting syndrome of swine, has two potential open reading frames, ORF1 and ORF2, greater than 600 nucleotides in length. ORF1 is predicted to encode a replication-associated protein (Rep) essential for replication of viral DNA, while ORF2 contains a conserved basic amino acid sequence at the N terminus resembling that of the major structural protein of chicken anaemia virus. Thus far, the structural protein(s) of PCV2 have not been identified. In this study, a viral structural protein of 30 kDa was identified in purified PCV2 particles. ORF2 of PCV2 was cloned into a baculovirus expression vector and the gene product was expressed in insect cells. The expressed ORF2 gene product had a molecular mass of 30 kDa, similar to that detected in purified virus particles. The recombinant ORF2 protein self-assembled to form capsid-like particles when viewed by electron microscopy. Antibodies against the ORF2 protein were detected in samples of sera obtained from pigs as early as 3 weeks after experimental infection with PCV2. These results show that the major structural protein of PCV2 is encoded by ORF2 and has a molecular mass of 30 kDa.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-81-9-2281
2000-09-01
2021-01-26
Loading full text...

Full text loading...

/deliver/fulltext/jgv/81/9/0812281a.html?itemId=/content/journal/jgv/10.1099/0022-1317-81-9-2281&mimeType=html&fmt=ahah

References

  1. Allan G. M., McNeilly F., Cassidy J. P., Reilly G. A. C., Adair B., Ellis W. A., McNulty M. S. 1995; Pathogenesis of porcine circovirus; experimental infections of colostrum deprived piglets and examination of pig foetal material. Veterinary Microbiology 44:49–64
    [Google Scholar]
  2. Allan G. M., McNeilly F., Kennedy S., Daft B., Clarke E. G., Ellis J. A., Haines D. M., Meehan B. M., Adair B. M. 1998; Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the USA and Europe. Journal of Veterinary Diagnostic Investigation 10:3–10
    [Google Scholar]
  3. An K., Gillock E. T., Sweat J. A., Reeves W. M., Consigli R. A. 1999; Use of the baculovirus system to assemble polyomavirus capsid-like particles with different polyomavirus structural proteins: analysis of the recombinant assembled capsid-like particles. Journal of General Virology 80:1009–1016
    [Google Scholar]
  4. Brown C. S., Van Lent J. W. M., Vlak J. M., Spaan W. J. M. 1991; Assembly of empty capsids by using baculovirus recombinants expressing human parvovirus B19 structural proteins. Journal of Virology 65:2702–2706
    [Google Scholar]
  5. Christensen J., Storgaard T., Bloch B., Alexandersen S., Aasted B. 1993; Expression of Aleutian mink disease parvovirus proteins in baculovirus vector system. Journal of Virology 67:229–238
    [Google Scholar]
  6. Clark E. G. 1997; Post-weaning multisystemic wasting syndrome. Proceedings of the American Association of Swine Practitioners, 28th Annual Meeting . Quebec City, Canada: pp 499–501
  7. Ellis J., Hassard L., Clark E., Harding J., Allan G., Willson P., Strokappe J., Martin K., McNeilly F., Meehan B., Todd D., Haines D. 1998; Isolation of circovirus from lesions of pigs with postweaning multisystemic wasting syndrome. Canadian Veterinary Journal 39:44–51
    [Google Scholar]
  8. Geissler K., Parrish C. R., Schneider K., Truyen U. 1999; Feline calicivirus capsid protein expression and self-assembly in cultured feline cells. Veterinary Microbiology 69:63–66
    [Google Scholar]
  9. Hamel A., Lin L. L., Nayar G. P. S. 1998; Nucleotide sequence of porcine circovirus associated with postweaning multisystemic wasting syndrome in pigs. Journal of Virology 72:5262–5267
    [Google Scholar]
  10. Harding J. C. 1997; Post-weaning multisystemic wasting syndrome (PMWS): preliminary epidemiology and clinical presentation. Proceedings of the American Association of Swine Practitioners, 28th Annual Meeting Quebec City, Canada: pp 503
    [Google Scholar]
  11. Harlow E., Lane D. 1988 Antibodies: A Laboratory Manual pp. 471–498 and 632–633 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  12. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  13. Lukert P., de Boer G. F., Dale J. L., Keese P., McNulty M. S., Randles J. W., Tischer I. 1995; The Circoviridae . In Virus Taxonomy . Sixth Report of the International Committee on Taxonomy of Viruses . pp 166–168 Edited by Murphy F. A., Fauquet C. M., Bishop D. H. L., Ghabrial S. A., Jarvis A. W., Martelli G. P., Mayo M. A., Summers M. D. Vienna & New York: Springer-Verlag;
  14. Mankertz A., Persson F., Mankertz J., Blaess G., Buhk H. 1997; Mapping and characterization of the origin of replication of porcine circovirus. Journal of Virology 71:2562–2566
    [Google Scholar]
  15. Mankertz A., Mankertz J., Wolf K., Buhk H.-J. 1998a; Identification of a protein essential for replication of porcine circovirus. Journal of General Virology 79:381–384
    [Google Scholar]
  16. Mankertz J., Buhk H., Blaess G., Mankertz A. 1998b; Transcription analysis of porcine circovirus (PCV). Virus Genes 16:267–276
    [Google Scholar]
  17. Meehan B. M., Creelan J. L., McNulty M. S., Todd D. 1997; Sequence of porcine circovirus DNA: affinities with plant circovirus. Journal of General Virology 78:221–227
    [Google Scholar]
  18. Meehan B. M., McNeilly F., Todd D., Kennedy S., Jewhurst V. A., Ellis J. A., Hassard L. E., Clark E. G., Haines D. M., Allan G. M. 1998; Characterization of novel circovirus DNAs associated with wasting syndromes in pigs. Journal of General Virology 79:2171–2179
    [Google Scholar]
  19. Morozov I., Sirinarumitr T., Sorden S. D., Halbur P. G., Morgan M. K., Yoon K., Paul P. S. 1998; Detection of a novel strain of porcine circovirus in pigs with postweaning multisystemic wasting syndrome. Journal of Clinical Microbiology 36:2535–2541
    [Google Scholar]
  20. O’Reilly D. R., Miller L. K., Luckow V. A. 1992 Baculovirus Expression Vectors: A Laboratory Manual pp 109–134 New York: W. H. Freeman;
  21. Ozawa K., Young N. 1987; Characterization of capsid and noncapsid protein of B19 parvovirus propagated in human erythroid bone marrow cell cultures. Journal of Virology 61:2627–2630
    [Google Scholar]
  22. Prasad B. V. V., Hardy M. E., Dokland T., Bella J., Rossmann M. G., Estes M. K. 1999; X-ray crystallographic structure of the Norwalk virus capsid. Science 286:287–290
    [Google Scholar]
  23. Ritchie B. W., Niagro F. D., Lukert P. D., Steffens W. L.III., Latimer K. S. 1989; Characterization of a new virus from cockatoos with psittacine beak and feather disease. Virology 171:83–88
    [Google Scholar]
  24. Shade R. O., Blundell M. C., Cotmore S. F., Tattersall P., Astell C. R. 1986; Nucleotide sequence and genome organization of human parvovirus B19 isolated from the serum of a child during aplastic crisis. Journal of Virology 58:921–936
    [Google Scholar]
  25. Sorden S. D., Harms P. A., Nawagitgul P., Cavanaugh D., Paul P. S. 1999; Development of a polyclonal-antibody-based immunohistochemical method for the detection of type 2 porcine circovirus in formalin-fixed, paraffin-embedded tissue. Journal of Veterinary Diagnostic Investigation 11:528–530
    [Google Scholar]
  26. Tischer I., Rasch R., Tochtermann G. 1974; Characterization of papovavirus- and picornavirus-like particles in permanent pig kidney cell lines. Zentralbatt fuer Bacteriologie Mikrobiologie und Hygiene Series A 226:153–167
    [Google Scholar]
  27. Tischer I., Gelderblom H., Vettermann W., Koch M. A. 1982; A very small porcine virus with circular single-stranded DNA. Nature 295:64–66
    [Google Scholar]
  28. Tischer I., Mields W., Wolff D., Vagt M., Griem W. 1986; Studies on epidemiology and pathogenicity of porcine circovirus. Archives of Virology 91:271–276
    [Google Scholar]
  29. Todd D., Creelan J. L., Mackie D. P., Rixon F., McNulty M. S. 1990; Purification and biochemical characterization of chicken anaemia agent. Journal of General Virology 71:819–823
    [Google Scholar]
  30. Wong S., Momoeda M., Field A., Kajigaya S., Young N. S. 1994; Formation of empty B19 parvovirus capsids by the truncated minor capsid protein. Journal of Virology 68:4690–4694
    [Google Scholar]
  31. Xing L., Kato K., Li T., Takeda N., Miyamura T., Hammar L., Cheng R. H. 1999; Recombinant hepatitis E capsid protein self-assembles into a dual-domain T =1 particle presenting native virus epitopes. Virology 265:35–45
    [Google Scholar]
  32. Zhang Y., Rameshwer D. S., Paul P. S. 1998; Monoclonal antibodies against conformationally dependent epitopes on porcine reproductive and respiratory syndrome virus. Veterinary Microbiology 63:125–136
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-81-9-2281
Loading
/content/journal/jgv/10.1099/0022-1317-81-9-2281
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error