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Nucleic Acids Research, 1994, Vol. 22, No. 2 208-213
© 1994

MOLECULAR BIOLOGY

Genetic identification and nucleotide sequence of the DNA polymerase gene of African swine fever virus

Alexandra Martins, Graça Ribeiro, M. Isabel Marques and JoãSo V. Costa*

Laboratory of Virology II, Gulbenkian Institute of Science Apartado 14, P-2781 Oeiras Codex, Portugal

*To whom correspondence should be addressed

Received September 23, 1993. Accepted December 17, 1993.

The DNA polymerase gene of African swine fever virus (ASFV) was mapped by marker rescue experiments using a phosphonoacetlc 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 aminoacid 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.


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