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Structure and expression inE. coli of the gene coding for protein p10 of African swine fever virus

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Summary

The gene encoding protein p10, a structural protein of African swine fever (ASF) virus, has been mapped, sequenced and expressed inE. coli. Protein p10 was purified from dissociated virus by reverse-phase HPLC, and its NH2-terminal end identified by automated Edman degradation. To map the gene encoding protein p10, a mixture of 20-mer oligonucleotides based upon a part of the amino acid sequence was hybridized to cloned ASF virus restriction fragments. This allowed the localization of the gene in fragmentEco RI K of the ASF virus genome. The nucleotide sequence obtained from this region revealed an open reading frame encoding 78 amino acids, with a high content of Ser and Lys residues. Several of the Ser residues are found in Ser-rich regions, which are also found in some nucleic acid-binding proteins. The gene coding for protein p10 has been inserted in an expression vector which contains the promoter for T 7 RNA polymerase. The recombinant plasmid was used to produce the ASF virus protein inE. coli. The bacterially produced p10 protein shows a strong DNA binding activity with similar affinity for both double-stranded and single-stranded DNA.

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Authors and Affiliations

  1. Departamento de Biología Funcional Facultad de Medicina, Universidad de Oviedo, Oviedo

    M. Muñoz, J. M. P. Freije & C. López-Otín

  2. Centro de Biología Molecular (CSIC-UAM), Facultad de Ciencias, Universidad Autónoma, Madrid, Spain

    M. L. Salas & E. Viñuela

Authors
  1. M. Muñoz
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  2. J. M. P. Freije
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  3. M. L. Salas
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  4. E. Viñuela
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  5. C. López-Otín
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Muñoz, M., Freije, J.M.P., Salas, M.L. et al. Structure and expression inE. coli of the gene coding for protein p10 of African swine fever virus. Archives of Virology 130, 93–107 (1993). https://doi.org/10.1007/BF01318999

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  • DOI: https://doi.org/10.1007/BF01318999

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