• 대한수의학회지
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• 제45권2호
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• pp.207-214
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• 2005

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.

• 대한수의학회지
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• 제42권1호
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• pp.89-100
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• 2002

The nonstructural glycoprotein NSP4, encoded by the 10th gene of rotavirus, has been known to play important roles in viral assembly and pathogenesis. The NSP4 genes of human rotavirus Korean isolates, designated as CBNU/HR-1, CBNU/HR-2, CBNU/HR-3, and CBNU/HR-4, were cloned, sequenced and characterized. Also, the NSP4 gene of the CBNU/HR-1 was expressed in a baculovirus-insect cell system. The sequence data indicated that the NSP4 genes of human rotavirus Korean isolates were 750 or 751 bases in length and encoded one open reading frame of 175 amino acids. Two glycosylation sites were recognized in the NSP4 gene of human rotavirus isolates tested. The NSP4 of CBNU/HR-1, CBNU/HR-3, and CBNU/HR-4 exhibited a high degree of amino acid sequence homology with that of NSP4 genotype B viruses, but a low degree of amino acid sequence homology with that of NSP4 genotype A viruses. However, the NSP4 of CBNU/HR-2 exhibited a high degree of amino acid sequence homology with that of NSP4 genotype A viruses, but a low degree of amino acid sequence homology with that of NSP4 genotype B viruses. The Sf9 cells infected with recombinant baculovirus, inserted with NSP4 gene of CBNU/HR-1, produced specific cytopathic effects and the expressed NSP4 was detected by immunofluorescence staining using NSP4-specific monoclonal antibody(MAb). The expressed NSP4 migrated at 16-26 kDa on SDS-PAGE and reacted with NSP4-specific MAb by Western blotting.

• 대한바이러스학회지
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• 제30권1호
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• pp.1-10
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• 2000

The nonstructural protein NSP4, encoded by gene 10 of rotavirus, has been shown to playa role in viral assembly and known to be an enterotoxin, causing diarrhea in mouse pups. NSP4 gene was cloned from CBNU-2 (virulent bovine rotavirus/diarrheic fecal sample) and CBNU-1 (cell-culture adapted bovine rotavirus/isolated from CBNU-2 and 75 times passaged on MA104 cells), respectively, by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequenced and compared. The sequence data indicated that the NSP4 genes of bovine rotavirus (BRV) were 751 bases in length and encoded one open reading frame of 175 amino acids beginning at base 42 and terminating at base 569. Differences in nucleotide sequence between CBNU-2 and CBNU-1 were observed at 6 positions (base 274, 296, 391, 394, 396 and 579). NSP4 gene of BRV exhibited a high degree of nucleotide (90% and 94%) and amino acid sequence (91% and 97%) homology with those of SA11 and UK but a low degree of nucleotide (77% and 79%) and amino acids sequence (81% and 85%) homology with those of Wa and OSU.

• 한국동물위생학회지
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• 제36권1호
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• pp.23-30
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• 2013

An avian rotavirus (AvRV-2) was isolated from feces of broilers suffering from acute gastroenteritis in 2011. It was the first avian rotavirus isolated in Korea. To investigate the molecular characteristics of AvRV-2, the VP4, VP6, VP7 and NSP4 gene nucleotide sequences were determined and compared with those of rotavirus strains available in the GenBank database. The phylogenetic tree of VP7 gene showed that AvRV-2 had a high degree of nucleotide sequence homology (93.4% to 94.7%) with those of rotaviruses belonging to genotype G19 cluster. The phylogenetic tree of the VP4 gene revealed a high degree of nucleotide sequence homology (95.8% to 95.9%) with genotype P[30] rotaviruses isolated from chickens. The VP6 and NSP4 gene nucleotide sequences showed the highest identities with those of avian strains with 95.3% to 96.4% and 90.3% to 92.2%, respectively. Genetic characterization of the VP4, VP6, VP7 and NSP4 showed that AvRV-2 strain was most closely related to chicken rotavirus strains from Germany and Japan. Comparative nucleotide sequences and phylogenetic analysis indicated that avian rotavirus isolated from broilers belonged to genotype G19P[30] and it was the first report on avian rotavirus infection in Korea.

• 대한수의학회지
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• 제48권4호
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• pp.423-429
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• 2008

Throughout the world, rotavirus infections cause extensive morbidity in human infants and diarrhea in animals such as white scour caused by bovine rotavirus in calves. We isolated three rotavirus strains designated KV0407, KV0418, and KV0426 from 103 fecal samples of diarrheic calves. The genes coding for proteins VP4, VP6, VP7, and NSP4 from strain KV0407 were sequenced and compared with the nucleotide sequences of other known strains of rotavirus. The KV0407 VP4 gene was highly homologous to the OSU (99.4%) and JL94 (99.4%), but not the B223 (62.4%) and K33 (62.4%) VP4 genes. The KV0407 and KV0418 VP7 genes were most similar to the OSU and super-short type VMRI VP7 genes. Based on nucleotide sequence analysis, the KV0407 strain was tentatively assigned to A serogroup (SG I), G5P[7], NSP4 genotype B and the KV0418 and KV0426 strains were assigned to A serogroup (SG I), G6P[5], NSP4 genotype A. The genetic characterization of these bovine rotavirus isolates could be useful for the diagnosis and prevention of diarrhea in calves.

• 한국동물위생학회지
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• 제35권2호
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• pp.83-90
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• 2012

Porcine rotaviruses are the most common causes of viral gastroenteritis in piglets around the world. The major G genotypes of porcine rotaviruses causing diarrhea were G4, G5 and G11 genotypes. Recently, G9 genotype rotaviruses were problemed at swine farms and frequently recognized from diarrheic piglets. In this study, a porcine rotavirus (PoRV-1) was isolated from piglet showing diarrhea using MA104 cells and confirmed as rotavirus by electron microscopy, genomic RNA electropherotyping and indirect immunofluorescence antibody tests. The nucleotide sequence of the VP7 gene of PoRV-1 was determined and compared with those of other genotype rotavirus strains from other parts of the world. Also, the nucleotide sequences of VP4, VP6 and NSP4 genes of PoRV-1 were determined and compared with those of other rotavirus strains from other countries. The results showed that the PoRV-1 isolate belonged to the G9 genotype and the P, I and E genotypes of PoRV-1 were P[23], I5 and E1, respectively. The Korean G9 PoRV-1 isolate and its nucleotide sequence data would be usefully used for the development of porcine rotavirus vaccines in near future.