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Mapping and sequence of the gene encoding the African swine fever virion protein of M_r 11500

Antonio Alcamí

Ana Angulo

Centro de Biología Molecular ‘Severo Ochoa’, Facultad de Ciencias, Universidad Autónoma, Cantoblanco, 28049 Madrid, Spain

The gene encoding the African swine fever virus protein of M_r 11500, present in the virus particle, has been mapped and sequenced in the genome of the Vero cell-adapted virus strain BA71V. A serum raised against virion proteins of M_r 12000 to 13000 isolated from polyacrylamide gels was used to screen a plasmid expression library, containing viral DNA random fragments, that expresses viral polypeptides fused to -galactosidase. Using this method, we have identified and sequenced the open reading frame (ORF) A137R, which initiates at the right end of the EcoRI A restriction fragment and extends into the EcoRI F fragment. Expression of the protein in Escherichia coli has confirmed that ORF A137R encodes a protein with an M_r of about 12000. A specific serum was raised against the E. coli-expressed protein, and has been used to identify the protein encoded by the ORF, which is translated at late times of infection and incorporated into the virus particle. Immunofluorescence experiments have shown that the protein localizes in virus factories.

Present address: Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, U.K.

Present address: Department of Immunology, The Scripps Research Institute, 10666 North Torrey Pines Road, La Jolla, California 92037, U.S.A.

Received 16 March 1993; accepted 30 June 1993.