title: Genetic identification and nucleotide sequence of the DNA polymerase gene of African swine fever virus.
abstract: The DNA polymerase gene of African swine fever virus (ASFV) was mapped by marker rescue experiments using a phosphonoacetic acid-resistant mutant and hybridization with an oligonucleotide probe designed from the most conserved motif of family B DNA polymerases. Viral DNA fragments mapping in this region were cloned and sequenced. An open reading frame coding for a 1244 amino acid long peptide with a molecular mass of 142.5 kDa was determined from the sequence. A unique feature of ASFV DNA polymerase is the presence of 13 tandem repeats of the sequence Ala-Gly-Asp-Pro near the carboxyl end of the molecule. Comparison with 30 sequences of alpha-like DNA polymerases of cellular and viral origin showed that ASFV DNA polymerase has all the conserved motifs of family B DNA polymerases. A 3.9 kb transcript was detected by Northern hybridization and the transcription initiation and termination sites were mapped by S1 analysis and primer extension. Late transcription was initiated at a site different from the early transcription initiation site. A 145 kDa protein, consistent with the size of the gene, was identified by an in situ enzyme assay after gel electrophoresis of infected cell extracts.
authors.affiliation: Laboratory of Virology II, Gulbenkian Institute of Science, Oeiras, Portugal.
authors.forename: A
authors.initials: A
authors.lastname: Martins
authors.forename: G
authors.initials: G
authors.lastname: Ribeiro
authors.forename: M I
authors.initials: MI
authors.lastname: Marques
authors.forename: J V
authors.initials: JV
authors.lastname: Costa
mesh_terms: D000358:African Swine Fever Virus / Q000201:enzymology / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D017124:Conserved Sequence; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic; D014709:Vero Cells
chemical_list: D004259:DNA-Directed DNA Polymerase