STRUCTURE AND EXPRESSION IN ESCHERICHIA-COLI OF THE GENE CODING FOR PROTEIN P10 OF AFRICAN SWINE FEVER VIRUS

The gene encoding protein p10, a structural protein of African swine f ever (ASF) virus, has been mapped, sequenced and expressed in E. 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 oligonucleot ides based upon a part of the amino acid sequence was hybridized to cl oned ASF virus restriction fragments. This allowed the localization of the gene in fragment Eco RI K of the ASF virus genome. The nucleotide sequence obtained from this region revealed an open reading frame enc oding 78 amino acids, with a high content of Ser and Lys residues. Sev eral of the Ser residues are found in Ser-rich regions, which are also found in some nucleic acid-binding proteins. The gene coding for prot ein p10 has been inserted in an expression vector which contains the p romoter for T7 RNA polymerase. The recombinant plasmid was used to pro duce the ASF virus protein in E. coli. The bacterially produced p10 pr otein shows a strong DNA binding activity with similar affinity for bo th double-stranded and single-stranded DNA.