Korean Journal of Veterinary Research (대한수의학회지)

The Korean Society of Veterinary Science (대한수의학회)
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Nucleotide sequence analysis and expression of NSP4 gene of human rotaviruses isolated in Korea

국내에서 분리된 사람 로타바이러스의 NSP4 유전자 염기서열 분석 및 발현

  • Jung, Dong-hyuk (Bioindist Co., Ltd.) ;
  • Song, Yun-kyung (Research Institute of Veterinary Medicine/College of Veterinary Medicine, Chungbuk National University) ;
  • Kim, Kyung-mi (Research Institute of Veterinary Medicine/College of Veterinary Medicine, Chungbuk National University) ;
  • Park, Hyo-sun (Research Institute of Veterinary Medicine/College of Veterinary Medicine, Chungbuk National University) ;
  • Back, Myoung-soon (Research Institute of Veterinary Medicine/College of Veterinary Medicine, Chungbuk National University) ;
  • Kang, Shien-young (Research Institute of Veterinary Medicine/College of Veterinary Medicine, Chungbuk National University)
  • 정동혁 ((주)바이오인디스트) ;
  • 송윤경 (충북대학교 수의과대학/동물의학연구소) ;
  • 김경미 (충북대학교 수의과대학/동물의학연구소) ;
  • 박효선 (충북대학교 수의과대학/동물의학연구소) ;
  • 백명순 (충북대학교 수의과대학/동물의학연구소) ;
  • 강신영 (충북대학교 수의과대학/동물의학연구소)
  • Accepted : 2002.03.02
  • Published : 2002.03.25
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Abstract

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.

Keywords

References

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