Cloning, characterization and expression of the gene that encodes the major neutralization-specific antigen of African horsesickness virus serotype 3.
The gene encoding the outer capsid protein, VP2, of African horsesickness virus serotype 3 (AHSV-3) has been sequenced in its entirety from cDNA clones of the segment 2 RNA, and compared with the previously published VP2 gene sequence of AHSV-4. AHSV-3 genome segment 2 was shown to be 3221 nucleotides in length, encoding a protein of 1057 amino acids with a 50.5% identity to the amino acid sequence of AHSV-4 VP2. Two areas of high variability (approximately 35% identity) were identified. The N-proximal variable region (amino acids 128 to 309) exhibited significant hydrophilicity, suggesting a possible role in the determination of the serotype-specific immune response. VP2 of AHSV-3 has furthermore been expressed in a baculovirus expression system. The expressed protein was shown to react specifically with an anti-AHSV-3 serum in Western blots. Antibodies raised in rabbits and guinea-pigs against the recombinant VP2 neutralized the virus in a plaque reduction assay, confirming the identity of VP2 as the major neutralization-specific antigen of AHSV.
Vreede
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Department of Genetics, University of Pretoria, South Africa.
Huismans
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D000356:African Horse Sickness Virus / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics*; D002213:Capsid / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D036022:Capsid Proteins; D003001:Cloning, Molecular; D015870:Gene Expression; D005814:Genes, Viral / Q000235:genetics*; D006168:Guinea Pigs; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009500:Neutralization Tests; D017924:Nucleopolyhedroviruses / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D012367:RNA, Viral / Q000096:biosynthesis; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D015678:Viral Structural Proteins / Q000235:genetics*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; C442519:VP2 protein, African horse sickness virus; D015678:Viral Structural Proteins