Comparative sequence analysis and expression of the M6 gene, encoding the outer capsid protein VP5, of African...

The entire nucleotide and deduced amino acid sequence of the M6 gene of African horsesickness virus (AHSV) serotype nine has been determined from four overlapping cDNA clones. The gene was found to be 1566 bp long, encoding a protein of 505 amino acids with a molecular weight of 56 737 Da and a nett charge of - 1 at neutral pH Comparative sequence analysis of the deduced amino acid sequence with the VP5 protein of AHSV-4, showed that only 81% of amino acids were conserved in type and position, although alternating regions of lower and higher conservation was identified by alignment of the primary sequences of different orbiviral VP5 proteins. Antigenically authentic AHSV-9 VP5 was also expressed in a baculovirus expression system and the expressed protein was shown to react specifically with anti-AHSV-9 as well as AHSV-3 serum in Western blot analysis.