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

Abstract

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.