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Expression in vivo and in vitro of the major structural protein (VP 73) of African swine fever virus

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Summary

The VP 73 protein was produced by in vitro transcription and translation from the Xho I-Bam HI fragment located between the Cla I-N and Cla I-H fragments of the viral genome. This DNA fragment encodes a late mRNA of about 2.6 kb detected in infected MS monkey and BHK hamster cells. The transcript was initiated at a site within two bases upstream of the translation initiation codon. The in vitro synthesized polypeptide shows the same molecular weight as the in vivo synthesized polypeptide, suggesting that VP 73 has no post-translational modification. There are two internal AUG initiation codons for in vitro translation, one of which is functional in vivo, as well as a possible GUG initiator codon detected by expression of the protein inE. coli cells.

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References

  1. Arzuza O (1990) Efecto de inhibidores de la sintesis de proteínas y DNA sobre la replicación del virus de la Peste Porcina Africana. Doctoral Tesis, Universidad Autónoma de Madrid, Spain

  2. Bertholet C, Stocco P, Van Meir E, Wittek R (1986) Functional analysis of the 5′ flanking sequence of a vaccinia late gene. EMBO J 5: 1951–1957

    Google Scholar 

  3. Blasco R, López-Otin C, Muñoz M, Bockamp E, Simon-Mateo C, Viñuela E (1990) Sequence and evolutionary relationships of African swine fever virus thymidine kinase. Virology 178: 301–304

    Google Scholar 

  4. Braun DK, Pereira L, Norrild B, Roizman B (1983) Application of denatured, electrophoretically separated, and immobilized lysates of herpes simplex virus-infected cells for the detection of monoclonal antibodies and for studies of the properties of viral proteins. J Virol 46: 103–112

    Google Scholar 

  5. Breese SS, DeBoer CJ (1966) Electron microscope observations of African swine fever virus in tissue culture cells. Virology 28: 420–428

    Google Scholar 

  6. Bucher Ph, Trifonov EN (1986) Compilation and analysis of eukaryotic POL II promoter sequences. Nucleic Acids Res 14: 10009–10026

    Google Scholar 

  7. Bujard H, Gentz R, Lanzer M, Stueber D, Mueller M, Ibrahimi I, Haeuptle MT, Dobberstein B (1987) A T 5 promoter-based transcription-translation system for the analysis of proteins in vitro and in vivo. Methods Enzymol 155: 416–433

    Google Scholar 

  8. Caballero RG, Tabarés E (1986) Application of pRPEL2 plasmid to detect African swine fever virus by DNA-DNA hybridization. Arch Virol 87: 119–125

    Google Scholar 

  9. Carvalho ZG, Rodrigues-Pousada C (1986) African swine fever virus gene expression in infected Vero cells. J Gen Virol 67: 1343–1350

    Google Scholar 

  10. Carrascosa AL, del Val M, Santaren JF, Viñuela E (1985) Purification and properties of African swine fever virus. J Virol 54: 337–344

    Google Scholar 

  11. Carrascosa JL, Carazo JM, Carrascosa AL, 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 

  12. Casal I, Enjuanes L, Viñuela E (1984) Porcine leukocyte cellular subsets sensitive to ASF virus in vitro. J Virol 52: 37–46

    Google Scholar 

  13. Davidson AJ, Moss B (1989) Structure of vaccinia virus late promoters. J Mol Biol 210: 771–784

    Google Scholar 

  14. Escribano JM, Tabarés E (1987) Proteins specified by African swine fever virus. V. Identification of immediate early, early and late proteins. Arch Virol 92: 221–232

    Google Scholar 

  15. García-Barreno B, Sanz A, Nogal ML, Viñuela E, Enjuanes L (1986) Monoclonal antibodies of African swine fever virus: antigenic differences among field virus isolates and viruses passaged in cell culture. J Virol 58: 385–392

    Google Scholar 

  16. Hanggi M, Bannwarth M, Stunnenberg HG (1986) Conserved TAAAT motif in vaccinia late promoters: overlapping TATA box and site transcription. EMBO J 5: 1071–1076

    Google Scholar 

  17. Hess WR (1971) African swine fever virus. In: Hess WR, Howell PG, Verwoerd DW (eds) African swine fever virus. Blue-tongue virus. Springer, Wien New York, pp 1–33 [Kingsbury DW, Zur Hausen H (eds) Virology monographs, vol 9]

    Google Scholar 

  18. Hess WR (1981) African swine fever: a reassessment. Adv Vet Sci Comp Med 25: 39–69

    Google Scholar 

  19. Kanda J, Melnick JL (1959) In vitro differentiation of virulent and attenuated polioviruses by their growth characteristics on MS cells. J Exp Med 109: 9–24

    Google Scholar 

  20. Letchworth GJ, Whyard TC (1984) Characterization of African swine fever virus antigenic proteins by immunoprecipitation. Arch Virol 80: 265–274

    Google Scholar 

  21. López-Otín C, Freije MP, Parra F, Mendez F, Viñuela E (1990) Mapping and sequence of the gene coding for protein p 72, the major capsid protein of African swine fever virus. Virology 175: 477–484

    Google Scholar 

  22. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor

    Google Scholar 

  23. Martín Hernández AM, Tabarés E (1991) Expression and characterization of the thymidine kinase gene of African swine fever virus. J Virol 65: 1046–1052

    Google Scholar 

  24. McCarthy JEG, Gualerzi C (1990) Translation control of prokaryotic gene expression. Trends Genet 6: 78–85

    Google Scholar 

  25. Mebus ChA (1988) African swine fever. Adv Virus Res 35: 251–269

    Google Scholar 

  26. Moura Nunes JF, Vigario JD, Terrina AM (1975) Ultrastructural study of African swine fever virus replication in cultures of swine bone marrow cells. Arch Virol 49: 59–66

    Google Scholar 

  27. Sánchez Botija C (1982) Peste Porcina Africana. Nuevos desarrollos. Rev Sci Tech Off Int Epic 1: 991–1029

    Google Scholar 

  28. Sanger F, Nickleen S, Coulson AR (1977) DNA sequencing with chain reactionterminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467

    Google Scholar 

  29. Sanz A, García-Barreno B, Nogal ML, Viñuela E, Enjuanes L (1985) Monoclonal antibodies specific for African swine fever virus proteins. J Virol 54: 199–206

    Google Scholar 

  30. Sargan DR, Gregory SP, Butterworth PHW (1982) A possible novel interaction between the 3′-end of 18 S ribosomal RNA and the 5′-leader sequence of many eukaryotic messengers RNAs. FEBS Lett 147: 133–136

    Google Scholar 

  31. Schloer GM (1985) Polypeptides and structure of African swine fever virus. Virus Res 3: 245–310

    Google Scholar 

  32. Tabarés E (1987) Detection of DNA viruses by radioactive and non-radioactive DNA probes: application to African swine fever virus. Arch Virol 92: 233–242

    Google Scholar 

  33. Tabarés E (1987) Characterization of African swine fever virus proteins. In: Becker Y (ed) African swine fever. Martinus Nijhoff, Boston, pp 51–61

    Google Scholar 

  34. Tabarés E, Marcotegui MA, Fernandez M, Sánchez Botija C (1980) Proteins specified by African swine fever virus. I. Analysis of the structural proteins and antigenic properties. Arch Virol 66: 107–117

    Google Scholar 

  35. Tabarés E, Martinez J, Gonzalvo FR, Sánchez Botija C (1980) Proteins specified by African swine fever virus. II. Analysis of proteins in infected cells and antigenic properties. Arch Virol 66: 119–132

    Google Scholar 

  36. Tabarés E, Fernández M, Salvador-Temprano E, Carnero ME, Sánchez Botija C (1981) A reliable enzyme linked immunosorbent assay for African swine fever using the major structural protein as antigenic reagent. Arch Virol 70: 297–300

    Google Scholar 

  37. Tabarés E, Martínez J, Martín E, Escribano JM (1983) Proteins specified by African swine fever virus. IV. Glycoproteins and phosphoproteins. Arch Virol 77: 167–180

    Google Scholar 

  38. Tabarés E, Olivares I, Santurde G, García MJ, Martín E, Carnero ME (1987) African swine fever virus DNA: deletions and additions during adaptation to growth in monkey kidney cells. Arch Virol 97: 333–346

    Google Scholar 

  39. Villinger F, Genovesi EV, Gerstner DJ, Whyard TC, Knudsen RC (1990) Inhibition of African swine fever virus in cultured swine monocytes by phosphonoacetic acid (PAA) and phosphonoformic acid (PFA). Arch Virol 115: 163–184

    Google Scholar 

  40. Viñuela E (1985) African swine fever virus. Curr Top Microbiol Immunol 116: 151–170

    Google Scholar 

  41. Wardley RC, Andrade CM, Black DN, Castro Portugal FL, Enjuanes L, Hess WR, Mebus Ch, Ordas A, Rutili D, Sánchez Vizcaino JM, Vigario JD, Wilkinson PJ (1983) African swine fever virus. Arch Virol 76: 73–90

    Google Scholar 

  42. Whyard TC, Wool SH, Letchworth GJ (1985) Monoclonal antibodies against African swine fever virus. Virology 142: 416–420

    Google Scholar 

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Cistué, C., Tabarés, E. Expression in vivo and in vitro of the major structural protein (VP 73) of African swine fever virus. Archives of Virology 123, 111–124 (1992). https://doi.org/10.1007/BF01317142

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