1887

Abstract

Bovine viral diarrhea virus (BVDV) belongs to the genus within the family . The lipid membrane of the virions is supposed to contain the three glycosylated envelope proteins E, E1 and E2, but detailed studies of virus assembly are complicated because no efficient purification method for pestiviruses has been described so far. In this study, we generated infectious BVDV with N-terminally FLAG-tagged E or E2 proteins, respectively. The expression of the epitope-tagged E and E2 proteins could be shown by immunofluorescence and Western blot experiments. Furthermore, an affinity tag purification protocol for the isolation and concentration of infectious BVDV was established. In the preparation with a titre of 10 TCID ml, spherical particles with a diameter of 43–58 nm (mean diameter: 48 nm) could be detected by negative staining electron microscopy, and immunogold labelling located both E and E2 proteins at the virus membrane.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.029330-0
2011-06-01
2019-10-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/92/6/1352.html?itemId=/content/journal/jgv/10.1099/vir.0.029330-0&mimeType=html&fmt=ahah

References

  1. Ahn B. C., Walz P. H., Kennedy G. A., Kapil S.. ( 2005;). Biotype, genotype, and clinical presentation associated with bovine viral diarrhea virus (BVDV) isolates from cattle. . Int J Appl Res Vet Med 3:, 319–325.
    [Google Scholar]
  2. Chu H. J., Zee Y. C.. ( 1984;). Morphology of bovine viral diarrhea virus. . Am J Vet Res 45:, 845–850.[PubMed]
    [Google Scholar]
  3. Collett M. S., Anderson D. K., Retzel E.. ( 1988;a). Comparisons of the pestivirus bovine viral diarrhea virus with members of the Flaviviridae. . J Gen Virol 69:, 2637–2643. [CrossRef][PubMed]
    [Google Scholar]
  4. Collett M. S., Larson R., Belzer S. K., Retzel E.. ( 1988;b). Proteins encoded by bovine viral diarrhea virus: the genomic organization of a pestivirus. . Virology 165:, 200–208. [CrossRef][PubMed]
    [Google Scholar]
  5. Coria M. F., Schmerr M. J., McClurkin A. W.. ( 1983;). Characterization of the major structural proteins of purified bovine viral diarrhea virus. . Arch Virol 76:, 335–339. [CrossRef][PubMed]
    [Google Scholar]
  6. Edwards S., Sands J. J., Harkness J. W.. ( 1988;). The application of monoclonal antibody panels to characterize pestivirus isolates from ruminants in Great Britain. . Arch Virol 102:, 197–206. [CrossRef][PubMed]
    [Google Scholar]
  7. Fauquet C. M., Mayo M. A., Maniloff J., Desselberger U., Ball L. A.. (editors) ( 2005;). Virus Taxonomy, VIIIth Report of the International Committee on Taxonomy of Viruses. London:: Elsevier/Academic Press;.
    [Google Scholar]
  8. Fetzer C., Tews B. A., Meyers G.. ( 2005;). The carboxy-terminal sequence of the pestivirus glycoprotein Erns represents an unusual type of membrane anchor. . J Virol 79:, 11901–11913. [CrossRef][PubMed]
    [Google Scholar]
  9. Gray E. W., Nettleton P. F.. ( 1987;). The ultrastructure of cell cultures infected with border disease and bovine virus diarrhoea viruses. . J Gen Virol 68:, 2339–2346. [CrossRef][PubMed]
    [Google Scholar]
  10. Holinka L. G., Fernandez-Sainz I., O’Donnell V., Prarat M. V., Gladue D. P., Lu Z., Risatti G. R., Borca M. V.. ( 2009;). Development of a live attenuated antigenic marker classical swine fever vaccine. . Virology 384:, 106–113. [CrossRef][PubMed]
    [Google Scholar]
  11. Houe H.. ( 1999;). Epidemiological features and economical importance of bovine virus diarrhoea virus (BVDV) infections. . Vet Microbiol 64:, 89–107. [CrossRef][PubMed]
    [Google Scholar]
  12. Iqbal M., Flick-Smith H., McCauley J. W.. ( 2000;). Interactions of bovine viral diarrhoea virus glycoprotein Erns with cell surface glycosaminoglycans. . J Gen Virol 81:, 451–459.[PubMed]
    [Google Scholar]
  13. Korn G.. ( 1977;). Zur Wirkung des Proteinase-Inhibitors Trasylol auf den Verlauf der Schweinepesterkrankung sowie zum Nachweis der Antigene (Virus und einer Chymotrypsin-like Protease) und der verschiedenen Antikörperarten. . Berl Munch Tierarztl Wochenschr 90:, 469–472 ( in German).[PubMed]
    [Google Scholar]
  14. Laird M. E., Desrosiers R. C.. ( 2007;). Infectivity and neutralization of simian immunodeficiency virus with FLAG epitope insertion in gp120 variable loops. . J Virol 81:, 10838–10848. [CrossRef][PubMed]
    [Google Scholar]
  15. Maurer K., Krey T., Moennig V., Thiel H. J., Rümenapf T.. ( 2004;). CD46 is a cellular receptor for bovine viral diarrhea virus. . J Virol 78:, 1792–1799. [CrossRef][PubMed]
    [Google Scholar]
  16. Merz A., Long G., Hiet M.-S., Brügger B., Chlanda P., Andre P., Wieland F., Krijnse-Locker J., Bartenschlager R.. ( 2011;). Biochemical and morphological properties of hepatitis C virus particles and determination of their lipidome. . J Biol Chem 286:, 3018–3032. [CrossRef][PubMed]
    [Google Scholar]
  17. Meyers G., Tautz N., Becher P., Thiel H. J., Kümmerer B. M.. ( 1996;). Recovery of cytopathogenic and noncytopathogenic bovine viral diarrhea viruses from cDNA constructs. . J Virol 70:, 8606–8613.[PubMed]
    [Google Scholar]
  18. Ohmann H. B.. ( 1990;). Electron microscopy of bovine virus diarrhoea virus. . Rev Sci Tech 9:, 61–73.[PubMed]
    [Google Scholar]
  19. Pestova T. V., Hellen C. U.. ( 1999;). Internal initiation of translation of bovine viral diarrhea virus RNA. . Virology 258:, 249–256. [CrossRef][PubMed]
    [Google Scholar]
  20. Peters W., Greiser-Wilke I., Moennig V., Liess B.. ( 1986;). Preliminary serological characterization of bovine viral diarrhoea virus strains using monoclonal antibodies. . Vet Microbiol 12:, 195–200. [CrossRef][PubMed]
    [Google Scholar]
  21. Poole T. L., Wang C., Popp R. A., Potgieter L. N., Siddiqui A., Collett M. S.. ( 1995;). Pestivirus translation initiation occurs by internal ribosome entry. . Virology 206:, 750–754. [CrossRef][PubMed]
    [Google Scholar]
  22. Pringle C. R.. ( 1998;). The universal system of virus taxonomy of the International Committee on Virus Taxonomy (ICTV), including new proposals ratified since publication of the Sixth ICTV Report in 1995. . Arch Virol 143:, 203–210. [CrossRef][PubMed]
    [Google Scholar]
  23. Riedel C., Lamp B., Heimann M., Rümenapf T.. ( 2010;). Characterization of essential domains and plasticity of the classical swine fever virus core protein. . J Virol 84:, 11523–11531. [CrossRef][PubMed]
    [Google Scholar]
  24. Ronecker S., Zimmer G., Herrler G., Greiser-Wilke I., Grummer B.. ( 2008;). Formation of bovine viral diarrhea virus E1-E2 heterodimers is essential for virus entry and depends on charged residues in the transmembrane domains. . J Gen Virol 89:, 2114–2121. [CrossRef][PubMed]
    [Google Scholar]
  25. Ross C. E., Dubovi E. J., Donis R. O.. ( 1986;). Herd problem of abortions and malformed calves attributed to bovine viral diarrhea. . J Am Vet Med Assoc 188:, 618–619.[PubMed]
    [Google Scholar]
  26. Rümenapf T., Unger G., Strauss J. H., Thiel H.-J.. ( 1993;). Processing of the envelope glycoproteins of pestiviruses. . J Virol 67:, 3288–3294.[PubMed]
    [Google Scholar]
  27. Rümenapf, T., Blome, S., Heimann, M. & Lamp, B. (2008). Core protein is not required for assembly of classical swine fever virus. In 7th ESVV Pestivirus Symposium.
  28. Takahashi H., Akazawa D., Kato T., Date T., Shirakura M., Nakamura N., Mochizuki H., Tanaka-Kaneko K., Sata T., Tanaka Y.. ( 2010;). Biological properties of purified recombinant HCV particles with an epitope-tagged envelope. . Biochem Biophys Res Commun 395:, 565–571. [CrossRef][PubMed]
    [Google Scholar]
  29. Thiel H. J., Stark R., Weiland E., Rümenapf T., Meyers G.. ( 1991;). Hog cholera virus: molecular composition of virions from a pestivirus. . J Virol 65:, 4705–4712.[PubMed]
    [Google Scholar]
  30. Unger T., Jacobovitch Y., Dantes A., Bernheim R., Peleg Y.. ( 2010;). Applications of the restriction free (RF) cloning procedure for molecular manipulations and protein expression. . J Struct Biol 172:, 34–44. [CrossRef][PubMed]
    [Google Scholar]
  31. Wegelt A., Reimann I., Zemke J., Beer M.. ( 2009;). New insights into processing of bovine viral diarrhea virus glycoproteins Erns and E1. . J Gen Virol 90:, 2462–2467. [CrossRef][PubMed]
    [Google Scholar]
  32. Weiland E., Stark R., Haas B., Rümenapf T., Meyers G., Thiel H.-J.. ( 1990;). Pestivirus glycoprotein which induces neutralizing antibodies forms part of a disulfide-linked heterodimer. . J Virol 64:, 3563–3569.[PubMed]
    [Google Scholar]
  33. Yu H., Grassmann C. W., Behrens S. E.. ( 1999;). Sequence and structural elements at the 3′ terminus of bovine viral diarrhea virus genomic RNA: functional role during RNA replication. . J Virol 73:, 3638–3648.[PubMed]
    [Google Scholar]
  34. Yu H., Isken O., Grassmann C. W., Behrens S. E.. ( 2000;). A stem-loop motif formed by the immediate 5′ terminus of the bovine viral diarrhea virus genome modulates translation as well as replication of the viral RNA. . J Virol 74:, 5825–5835. [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.029330-0
Loading
/content/journal/jgv/10.1099/vir.0.029330-0
Loading

Data & Media loading...

Supplements

vol. , part 6, pp. 1352 - 1357

Sequences of primers used in this study [PDF](36 KB)



PDF

Most Cited This Month

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