title: A first full outer capsid protein sequence data-set in the Orbivirus genus (family Reoviridae): cloning, sequencing, expression and analysis of a complete set of full-length outer capsid VP2 genes of the nine African horsesickness virus serotypes.
abstract: The outer capsid protein VP2 of African horsesickness virus (AHSV) is a major protective antigen. We have cloned full-length VP2 genes from the reference strains of each of the nine AHSV serotypes. Baculovirus recombinants expressing the cloned VP2 genes of serotypes 1, 2, 4, 6, 7 and 8 were constructed, confirming that they all have full open reading frames. This work completes the cloning and expression of the first full set of AHSV VP2 genes. The clones of VP2 genes of serotypes 1, 2, 5, 7 and 8 were sequenced and their amino acid sequences were deduced. Our sequencing data, together with that of the published VP2 genes of serotypes 3, 4, 6 and 9, were used to generate the first complete sequence analysis of all the (sero)types for a species of the Orbivirus genus. Multiple alignment of the VP2 protein sequences showed that homology between all nine AHSV serotypes varied between 47.6 % and 71.4 %, indicating that VP2 is the most variable AHSV protein. Phylogenetic analysis grouped together the AHSV VP2s of serotypes that cross-react serologically. Low identity between serotypes was demonstrated for specific regions within the VP2 amino acid sequences that have been shown to be antigenic and play a role in virus neutralization. The data presented here impact on the development of new vaccines, the identification and characterization of antigenic regions, the development of more rapid molecular methods for serotype identification and the generation of comprehensive databases to support the diagnosis, epidemiology and surveillance of AHS.
authors.affiliation: Onderstepoort Veterinary Institute, 0110 Onderstepoort, South Africa.
authors.forename: A C
authors.initials: AC
authors.lastname: Potgieter
authors.affiliation: Onderstepoort Veterinary Institute, 0110 Onderstepoort, South Africa.
authors.forename: M
authors.initials: M
authors.lastname: Cloete
authors.affiliation: Division of Biochemistry, School for Chemistry and Biochemistry, University of Potchefstroom for CHE, 2520 Potchefstroom, South Africa.
authors.forename: P J
authors.initials: PJ
authors.lastname: Pretorius
authors.affiliation: Onderstepoort Veterinary Institute, 0110 Onderstepoort, South Africa.
authors.forename: A A
authors.initials: AA
authors.lastname: van Dijk
mesh_terms: D000355:African Horse Sickness / Q000821:virology; D000356:African Horse Sickness Virus / Q000145:classification* / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D006736:Horses; D051379:Mice; D008969:Molecular Sequence Data; D018163:Orbivirus / Q000145:classification* / Q000235:genetics; D010802:Phylogeny; D011995:Recombination, Genetic; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012703:Serotyping
chemical_list: D036022:Capsid Proteins; C442519:VP2 protein, African horse sickness virus