1887

Abstract

The genomic RNA of a porcine reproductive and respiratory syndrome virus (PRRSV) isolate from the U.S.A., VR 2385 (ATCC), was copied into cDNA after priming with oligo(dT) and cloned into phage lambda. The cDNA clones representing the 3′-terminal genomic RNA of the virus were isolated and sequenced. The genome is a positive-stranded, polyadenylated RNA with an estimated size of 15 kb. Analysis of the resulting sequence identified three complete open reading frames (ORFs) with the potential to encode polypeptides with predicted s of 22·2K (ORF 5), 19·1K (ORF 6) and 13·6K (ORF 7). ORF 7, which is closest to the 3′ end, is predicted to encode a highly basic nucleocapsid protein displaying 58 % amino acid identity to the corresponding protein of the Lelystad virus (LV), a European PRRSV isolate. ORFs 6 and 5, preceding ORF 7, are each predicted to encode proteins containing several hydrophobic domains that are thought to be membrane- associated. The VR 2385 ORF 6 protein is the most conserved structural protein. It has 78% amino acid identity to the equivalent LV protein, and ORF 5 shares only 54% of its amino acid sequence. Northern blot analysis revealed a 3′-coterminal nested set of six subgenomic RNAs in VR 2385 virus-infected CRL 11171 cells. Our results indicate that VR 2385, like LV, is a member of the newly proposed arterivirus group. However, the striking genetic variation and the difference in pathogenicity between LV and VR 2385 suggest that the viruses causing PRRS in the U.S.A. and Europe are highly variable and they may represent different genotypes.

The nucleotide sequence data reported in this paper have been deposited with the GenBank database and assigned the accession number U03040.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-75-7-1795
1994-07-01
2021-10-18
Loading full text...

Full text loading...

/deliver/fulltext/jgv/75/7/JV0750071795.html?itemId=/content/journal/jgv/10.1099/0022-1317-75-7-1795&mimeType=html&fmt=ahah

References

  1. Bautista E. M., Goyal S. M., Yoon I. J., Joo H. S., Collins J. E. 1993; Comparison of porcine alveolar macrophages and CL 2621 for the detection of porcine reproductive and respiratory syndrome (PRRS) virus and anti-PRRS antibody. Journal of Veterinary Diagnostic Investigation 5:163–165
    [Google Scholar]
  2. Benfield D. A., Nelson E., Collins J. E., Harris L., Goyal S. M., Robison D., Christianson W. T., Morrison R. B., Gorcyca D., Chladek D. 1992; Characterization of swine infertility and respiratory syndrome (SIRS) virus (isolate ATCC VR-2332). Journal of Veterinary Diagnostic Investigation 4:127–133
    [Google Scholar]
  3. Chen Z., Kuo L., Rowland R. R. R., Even C., Faaberg K. S., Plagemann P. G. W. 1993; Sequences of 3′ end of genome and of 5′ end of open reading frame la of lactate dehydrogenase-elevating virus and common junction motifs between 5′ leader and bodies of seven subgenomic mRNAs. Journal of General Virology 74:643–660
    [Google Scholar]
  4. Christianson W. T., Collins J. E., Benfield D. A., Harris L., Gorcyca D. E., Chladek D. W., Morrison R. B., Joo H. S. 1992; Experimental reproduction of swine infertility and respiratory syndrome in pregnant sows. American Journal of Veterinary Research 53:485–488
    [Google Scholar]
  5. Collins J. E., Benfield D. A., Christianson W. T., Harris L., Hennings J. C., Shaw D. P., Goyal S. M., Mccullough S., Morrison R. B., Joo H. S., Gorcyca D. E., Chladek D. W. 1992; Isolation of swine infertility and respiratory syndrome virus (isolate ATCC VR-2332) in North America and experimental reproduction of the disease in gnotobiotic pigs. Journal of Veterinary Diagnostic Investigation 4:117–126
    [Google Scholar]
  6. Conzelmann K.-K., Visser N., Van Woensel P., Thiel H.-J. 1993; Molecular characterization of porcine reproductive and respiratory syndrome virus, a member of the arterivirus group. Virology 193:329–339
    [Google Scholar]
  7. Dea S., Bilodeau R., Athanaseous R., Sauvageau R. A., Martineau G. P. 1992; PRRS syndrome in Quebec: isolation of a virus serologically related to Lelystad virus. Veterinary Record 130:167
    [Google Scholar]
  8. Den Boon J. A., Snijder E. J., Chirnside E. D., De Vries A. A. F., Horzinek M. C., Spaan W. J. M. 1991; Equine arteritis virus is not a togavirus but belongs to the coronavirus-like superfamily. Journal of Virology 65:2910–2920
    [Google Scholar]
  9. De Vries A. A. F., Chirnside E. D., Bredenbeek P. J., Gravestein L. A., Horzinek M. C., Spaan W. J. M. 1990; All subgenomic mRNAs of equine arteritis virus contain a common leader sequence. Nucleic Acids Research 18:3241–3247
    [Google Scholar]
  10. Godeny E. K., Chen L., Kumar S. N., Methven S. L., Koonin E. V., Brinton M. A. 1993; Complete genomic sequence and phylogenetic analysis of the lactate dehydrogenase-elevating virus (LDV). Virology 194:585–596
    [Google Scholar]
  11. Hill H. 1990; Overview and history of mystery swine disease (swine infertility and respiratory syndrome). In Proceedings of the Mystery Swine Disease Committee Meeting, Denver, Colorado pp. 29–31 Denver: Livestock Conservation Institute;
    [Google Scholar]
  12. Kapke P. A., Brian D. A. 1986; Sequence analysis of the porcine transmissible gastroenteritis coronavirus nucleocapsid protein gene. Virology 151:41–49
    [Google Scholar]
  13. Keffaber K. K. 1989; Reproductive failure of unknown etiology. American Association of Swine Practitioners Newsletter 1:1–9
    [Google Scholar]
  14. Kuo L., Harty J. T., Erickson L., Palmer G. A., Plagemann P. G. W. 1991; A nested set of eight RNAs is formed in macrophages infected with lactate dehydrogenase-elevating virus. Journal of Virology 65:5118–5123
    [Google Scholar]
  15. Kuo L., Chen Z., Rowland R. R. R., Faaberg K. S., Plagemann P. G. W. 1992; Lactate dehydrogenase-elevating virus (LDV): subgenomic mRNAs, mRNA leader and comparison of 3′-terminal sequences of two LDV isolates. Virus Research 23:55–72
    [Google Scholar]
  16. Loula T. 1991; Mystery pig disease: an update for the practitioner. Agri-Practice 12:23–34
    [Google Scholar]
  17. Meng X.-J., Paul P. S., Vaughn E. M., Zimmerman J. J. 1993; Development of a radiolabeled nucleic acid probe for the detection of encephalomyocarditis virus of swine. Journal of Veterinary Diagnostic Investigation 5:254–258
    [Google Scholar]
  18. Meulenberg J. J. M., Hulst M. M., De Meijer E. J., Moonen P. L. J. M., Den Besten A., De Kluyver E. P., Wensvoort G., Moormann R. J. M. 1993a; Lelystad virus, the causative agent of porcine epidemic abortion and respiratory syndrome (PEARS), is related to LDV and EAV. Virology 192:62–72
    [Google Scholar]
  19. Meulenberg J. J. M., De Meijer E. J., Moormann R. J. M. 1993b; Subgenomic RNAs of Lelystadvirus contain a conserved leader-body junction sequence. Journal of General Virology 74:1697–1701
    [Google Scholar]
  20. Nelson E. A., Christopher-Hennings J., Drew T., Wensvoort G., Collins J. E., Benfield D. A. 1993; Differentiation of U.S. and European isolates of porcine reproductive and respiratory syndrome virus by monoclonal antibodies. Journal of Clinical Microbiology 31:3184–3189
    [Google Scholar]
  21. Paton D. J., Brown I. H., Edwards S., Wensvoort G. 1991; Blue ear disease of pigs. Veterinary Record 128:617
    [Google Scholar]
  22. Paul P. S., Halbur P. G., Meng X.-J. 1993; Porcine reproductive and respiratory syndrome: an overview. Journal of Clinical Veterinary Medicine 11:19–28
    [Google Scholar]
  23. Plagemann P. G. W., Moennig V. 1992; Lactate dehydrogenase-elevating virus, equine arteritis virus and simian hemorrhagic fever virus, a new group of positive strand RNA virus. Advances in Virus Research 41:99–192
    [Google Scholar]
  24. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual, 2nd edn. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  25. Spaan W., Cavanagh D., Horzinek M. C. 1988; Coronaviruses: structure and genome expression. Journal of General Virology 69:2939–2952
    [Google Scholar]
  26. Strauss J. H., Strauss E. G. 1988; Evolution of RNA viruses. Annual Review of Biochemistry 42:657–683
    [Google Scholar]
  27. Terpstra C., Wensvoort G., Pol J. M. A. 1991; Experimental reproduction of porcine epidemic abortion and respiratory syndrome (mystery swine disease) by infection with Lelystad virus: Koch’s postulates fulfilled. Veterinary Quarterly 13:131–136
    [Google Scholar]
  28. Wensvoort G. 1993; Lelystad virus and the porcine epidemic abortion and respiratory syndrome. Veterinary Research 24:117–124
    [Google Scholar]
  29. Wensvoort G., Terpstra C., Pol J. M., Ter Laak E. A., Bloemraad M., de Kluyver E. P., Kragten C., van Buiten L., den Besten A., Wagenaar F., Broekhuusen J. M., Moonen P. L. J. M., Zetstra T., de Boer E. A., Tibben H. J., de Jong M. F., Van’t Veld P., Groenland G. J. R., van Gennep J. A., Voets M. T., Verheijden J. H. M., Braamskamp J. 1991; Mystery swine disease in the Netherlands: the isolation of Lelystad virus. Veterinary Quarterly 13:121–130
    [Google Scholar]
  30. Wensvoort G., de Kluyver E. P., Luijtze E. A., den Besten A., Harris L., Collins J. E., Christianson W. T., Chladek D. 1992; Antigenic comparison of Lelystad virus and swine infertility and respiratory syndrome virus. Journal of Veterinary Diagnostic Investigation 4:134–138
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-75-7-1795
Loading
/content/journal/jgv/10.1099/0022-1317-75-7-1795
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