Nucleotide sequence analysis and expression of NSP4 gene of avian rotavirus |
In-ho Shin1, Seung-chul Lee1, Won-yong Kim2, Shien-young Kang1 |
1Research Institute of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University 2College of Medicine, Chungang University |
조류 로타바이러스의 NSP4 유전자 염기서열분석 및 발현 |
신인호1, 이승철1, 김원용2, 강신영1 |
1충북대학교 수의과대학 동물의학연구소 2중앙대학교 의과대학 |
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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. |
Key Words:
avian rotavirus, NSP4, sequence, expression |
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