| Nucleotide sequence analysis and expression of NSP4 gene of human rotaviruses isolated in Korea |
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Dong-hyuk Jung1, Yun-kyung Song2, Kyung-mi Kim2, Hyo-sun Park2, Myoung-soon Back2, Shien-young Kang2 |
1Bioindist Co., Ltd.
2Research Institute of Veterinary Medicine/College of Veterinary Medicine, Chungbuk National University |
| 국내에서 분리된 사람 로타바이러스의 NSP4 유전자 염기서열 분석 및 발현 |
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정동혁1, 송윤경2, 김경미2, 박효선2, 백명순2, 강신영2 |
1(주)바이오인디스트
2충북대학교 수의과대학/동물의학연구소 |
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| Abstract |
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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. |
| Key Words:
Human rotavirus, NSP4, nucleotide sequence, baculovirus, expression |
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