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Nucleotide sequence analysis and expression of NSP4 gene of avian rotavirus  

Shin, In-ho (Research Institute of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University)
Lee, Seung-chul (Research Institute of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University)
Kim, Won-yong (College of Medicine, Chungang University)
Kang, Shien-young (Research Institute of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University)
Publication Information
Korean Journal of Veterinary Research / v.45, no.2, 2005 , pp. 207-214 More about this Journal
Abstract
The nonstructural protein 4 (NSP4) of rotavirus encoded by gene 10, plays an important role in rotavirus pathogenicity. In this study, NSP4 gene of avian rotavirus (AvRV-1, AvRV-2) was analyzed and expressed using baculovirus expression system. The sequence data indicated that the NSP4 gene of AvRV-1 and AvRV-2 were 727 bases in length, encoded one open reading frame of 169 amino acids beginning at base 41 and terminating at base 550, and had two glycosylation sites. Nucleotide sequences of NSP4 gene of AvRV-1 and AvRV-2 exhibited a high degree of homology ($88.1{\pm}7.6%$) with avian rotaviruses, namely Ty1, Ty3 and PO-13. Phylogenetic analysis showed that AvRV-1 and AvRV-2 belonged to genotype NSP4[E], which is widely found in group A avian rotaviruses. The baculovirus-expressed NSP4 migrated at 20-28 kDa and reacted with NSP4-specific antiserum by FA and Western blot. Furthermore, it was found to be a glycoprotein by using tunicamycin, which is a specific inhibitor of N-linked glycosylation.
Keywords
avian rotavirus; NSP4; sequence; expression;
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