Mapping and sequence of the gene encoding the African swine fever virion protein of 11500 Free

Abstract

The gene encoding the African swine fever virus protein of 11500, present in the virus particle, has been mapped and sequenced in the genome of the Vero cell-adapted virus strain BA71V. A serum raised against virion proteins of 12000 to 13000 isolated from polyacrylamide gels was used to screen a plasmid expression library, containing viral DNA random fragments, that expresses viral polypeptides fused to -galactosidase. Using this method, we have identified and sequenced the open reading frame (ORF) A137R, which initiates at the right end of the RI A restriction fragment and extends into the RI F fragment. Expression of the protein in has confirmed that ORF A137R encodes a protein with an of about 12000. A specific serum was raised against the -expressed protein, and has been used to identify the protein encoded by the ORF, which is translated at late times of infection and incorporated into the virus particle. Immunofluorescence experiments have shown that the protein localizes in virus factories.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-74-11-2317
1993-11-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/74/11/JV0740112317.html?itemId=/content/journal/jgv/10.1099/0022-1317-74-11-2317&mimeType=html&fmt=ahah

References

  1. Aguado B., Viñuela E., Alcamí A. 1991; African swine fever virus fatty acid acylated proteins. Virology 185:942–945
    [Google Scholar]
  2. Alcamí A., Angulo A., López-Otín C., Muñoz M., Freije J. M. P., Carrascosa A. L., Viñuela E. 1992; Amino acid sequence and structural properties of protein pi2, an African swine fever virus attachment protein. Journal of Virology 66:3860–3868
    [Google Scholar]
  3. Almazán F., Rodríguez J. M., Andrés G., Pérez R., Viñuela E., Rodríguez J. F. 1992; Transcriptional analysis of multigene family 110 of African swine fever virus. Journal of Virology 66:6655–6667
    [Google Scholar]
  4. Almazán F., Rodríguez J. M., Angulo A., Viñuela E., Rodríguez J. F. 1993; Transcriptional mapping of a late gene coding for the p12 attachment protein of African swine fever virus. Journal of Virology 67:553–556
    [Google Scholar]
  5. Angulo A., Viñuela E., Alcamí A. 1993; Inhibition of African swine fever virus binding and infectivity by purified, recombinant virus attachment protein pl2. Journal of Virology 67:5463–5476
    [Google Scholar]
  6. Bonner W. M., Laskey R. A. 1974; A film detection method for tritium-labeled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46:83–88
    [Google Scholar]
  7. Camacho A., Viñuela E. 1991; Protein p22 of African swine fever virus: an early structural protein that is incorporated into the membrane of infected cells. Virology 181:251–257
    [Google Scholar]
  8. Carrascosa A. L., Del Val M., Santarén J. F., Viñuela E. 1985; Purification and properties of African swine fever virus. Journal of Virology 54:337–344
    [Google Scholar]
  9. Carrascosa A. L., Sastre I., Viñuela E. 1991; African swine fever virus attachment protein. Journal of Virology 65:2283–2289
    [Google Scholar]
  10. Carrascosa J. L., Carazo J. M., Carrascosa A. L., García N., Santisteban A., Viñuela E. 1984; General morphology and capsid fine structure of African swine fever virus particles. Virology 132:160–172
    [Google Scholar]
  11. Costa J. 1990; African swine fever virus. In Molecular Biology of Iridoviruses pp 247–270 Edited by Darai G. Boston: Kluwer Academic Publishers;
    [Google Scholar]
  12. De Boer C. J. 1967; Studies to determine neutralizing antibodies in sera from animals recovered from African swine fever and laboratory animals inoculated with African virus with adjuvants. Archiv fur die gesamte Virusforschung 20:164–179
    [Google Scholar]
  13. Del Val M., Viñuela E. 1987; Glycosylated components induced in African swine fever (ASF) virus-infected Vero cells. Virus Research 7:297–308
    [Google Scholar]
  14. Del Val M., Carrascosa J. L., Viñuela E. 1986; Glycosylated components of African swine fever virus. Virology 152:39–49
    [Google Scholar]
  15. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Research 12:387–395
    [Google Scholar]
  16. Dixon L. K., Wilkinson P. J., Sumption K. J., Ekue F. 1990; Diversity of the African swine fever virus genome. In Molecular Biology of Iridoviruses pp 271–295 Edited by Darai G. Boston: Kluwer Academic Publishers;
    [Google Scholar]
  17. Esteves A., Marques M. I., Costa J. V. 1986; Two-dimensional analysis of African swine fever proteins and proteins induced in infected cells. Virology 152:181–191
    [Google Scholar]
  18. García-Barreno B., Sanz A., Nogal M. L., Viñuela E., Enjuanes L. 1986; Monoclonal antibodies of African swine fever virus: antigenic differences among field virus isolates and viruses passaged in cell culture. Journal of Virology 58:385–392
    [Google Scholar]
  19. Harlow E., Lane D. 1988Antibodies: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
  20. Kuznar J., Salas M. L., Viñuela E. 1980; DNA-dependent RNA polymerase in African swine fever virus. Virology 101:169–175
    [Google Scholar]
  21. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  22. Ley V., Almendral J. M., Carbonero P., Beloso A., Viñuela E., Talavera A. 1984; Molecular cloning of African swine fever virus DNA. Virology 133:249–257
    [Google Scholar]
  23. López-Otín C., Simón C., Méndez E., Viñuela E. 1988; Mapping and sequence of the gene encoding protein p37, a major structural protein of African swine fever virus. Virus Genes 1:291–303
    [Google Scholar]
  24. López-Otín C., Simón-Mateo C., Martínez L., Viñuela E. 1989; Gly-Gly-X, a novel consensus sequence for the proteolytic processing of viral and cellular proteins. Journal of Biological Chemistry 264:9107–9110
    [Google Scholar]
  25. López-Otín C., Freije J. M. P., Parra F., Méndez E., Viñuela E. 1990; Mapping and sequence of the gene coding for protein p72, the major capsid protein of African swine fever virus. Virology 175:477–484
    [Google Scholar]
  26. Rodríguez J. M., Almazán F., Viñuela E., Rodríguez J. F. 1992; Genetic manipulation of African swine fever virus: construction of recombinant viruses expressing the β-galactosidase gene. Virology 188:67–76
    [Google Scholar]
  27. Salas M. L., Kuznar J., Viñuela E. 1981; Polyadenylation, methylation, and capping of the RNA synthesized in vitro by African swine fever virus. Virology 113:484–491
    [Google Scholar]
  28. Sambrook J., Fritsch E. F., Maniatis T. 1989Molecular Cloning: A Laboratory Manual 2nd edn New York: Cold Spring Harbor Laboratory;
  29. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, U,. S,. A 74:5463–5467
    [Google Scholar]
  30. Santarén J. F., Viñuela E. 1986; African swine fever virus-induced polypeptides in VERO cells. Virus Research 5:391–105
    [Google Scholar]
  31. Sanz A., García-Barreno B., Nogal M. L., Viñuela E., Enjuanes L. 1985; Monoclonal antibodies specific for African swine fever virus proteins. Journal of Virology 54:199–206
    [Google Scholar]
  32. Simón-Mateo C., Andrés G., Viñuela E. 1993; Polyprotein processing in African swine fever virus: a novel strategy of gene expression for a DNA virus. EMBO Journal 12:2977–2987
    [Google Scholar]
  33. Stanley K. K. 1983; Solubilization and immune detection of β-galactosidase hybrid proteins carrying foreign antigenic determinants. Nucleic Acids Research 11:4077–4092
    [Google Scholar]
  34. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. 1990; Use of T7 RNA polymerase to direct expression of cloned genes. Methods in Enzymology 185:60–89
    [Google Scholar]
  35. Vieira J., Messing J. 1982; The pUC plasmids, an M13mp7-derived system for insertion, mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
    [Google Scholar]
  36. Viñuela E. 1985; African swine fever virus. Current Topics in Microbiology and Immunology 116:155–170
    [Google Scholar]
  37. Viñuela E. 1987; Molecular biology of African swine fever virus. In African Swine Fever pp 31–49 Edited by Becker Y. Boston: Martinus Nijhoff Publishing;
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-74-11-2317
Loading
/content/journal/jgv/10.1099/0022-1317-74-11-2317
Loading

Data & Media loading...

Most cited Most Cited RSS feed