Amino Acid Tandem Repeats within a Late Viral Gene Define the Central Variable Region of African Swine Fever Virus

AbstractThe central variable region (CVR) of the African swine fever virus (ASFV) genome is contained within the 9-RL open reading frame (ORF). ORF 9-RL of the ASFV isolate Malawi Lil-20/1 predicts a protein of 614 amino acids with amino- and carboxy-terminal hydrophobic regions and a centrally located hydrophilic region. The CVR of the genome, located centrally within this ORF, is 372 bp and contains a 132-bp direct repeat. The translated CVR within ORF 9-RL contains 31 tandem tetramers, predominantly NADT, NANT, NVDT, and, in a few cases, CAST, GAST, or CADT.In vitrotranslation of 9-RL yielded a 94-kDa protein that was strongly reactive with convalescent pig serum while monospecific 9-RL antiserum identified a late viral protein of 94 kDa in ASFV-infected macrophages. The protein, detected by immunofluorescence staining with 9-RL antiserum, was distributed homogeneously throughout the cytoplasm of infected Vero cells. 9-RL protein size varied among different viral isolates and among cell-culture-adapted viruses. Protein size increased proportionately with the degree of cell culture adaptation and was directly correlated with the size of the CVR present within the ORF (300–500 bp). Analysis of the number and composition of tandem tetramers present within the CVR of a given ASFV isolate may prove useful for identifying and/or grouping ASFV isolates