Molecular Characterization of a Korean Isolate of Human Norovirus, the Hu/NLV/Gunpo/2006/KO Strain

인체 노로바이러스의 한국분리주 Hu/NLV/Gunpo/2006/KO의 분자생물학적 특성

  • Jeong, Ah-Yong (Department of Microbiology, College of Medicine and Medical Research Institute, Chungbuk National University) ;
  • Yun, Sang-Im (Department of Microbiology, College of Medicine and Medical Research Institute, Chungbuk National University) ;
  • Jee, Young-Mee (Division of Viral Hepatitis and Poliovirus, Center for Infectious Diseases, National Institute of Health) ;
  • Kang, Yoon-Sung (Department of Bio-Material, DNA Repair Research Center, Graduate School, Chosun University) ;
  • Lee, Young-Min (Department of Microbiology, College of Medicine and Medical Research Institute, Chungbuk National University)
  • 정아용 (충북대학교 의과대학 미생물학교실, 기초의학연구소) ;
  • 윤상임 (충북대학교 의과대학 미생물학교실, 기초의학연구소) ;
  • 지영미 (질병관리본부 국립보건연구원 감염병센터 간염폴리오바이러스과) ;
  • 강윤성 (조선대학교 생물신소재학과) ;
  • 이영민 (충북대학교 의과대학 미생물학교실, 기초의학연구소)
  • Received : 2009.05.11
  • Accepted : 2009.06.08
  • Published : 2009.06.30

Abstract

Norovirus (NV) with a variety of genotypes, a member of the family Caliciviridae, causes acute nonbacterial gastroenteritis in humans. We determined the nucleotide sequence of three open reading frames (ORFs) of a NV Korean strain and characterized the genetic relationship with others. The Korean strain designated Hu/NLV/Gunpo/2006/KO was isolated from the stool specimen of a 2-year-old female suffering from gastroenteritis. By performing reverse transcription and PCR amplification, three overlapping cDNAs were synthesized and used for direct sequencing. We found that like other NVs, this strain contains three ORFs: ORF1, 5,100 bp; ORF2, 1,647 bp; ORF3, 765 bp. Of 35 NVs, ORF1 had a level of genetic diversity lower than ORF2 and ORF3, of which the C-termini of the ORF2 and ORF3 showed a relatively high degree of genetic diversity. Phylogenetic analyses indicated that the Korean strain belonged to genogroup II, with Saitama U1, Gifu'96, Mc37, and Vietnam 026 being formed a single genetic cluster. The nucleotide sequence information of three ORFs of a NV Korean isolate will be useful not only for the development of a diagnostic tool and understanding of genetic relationship, but also provide important basic information for the functional analysis of their gene products.

노로바이러스는 급성 위장염을 일으키는 Caliciviridae 과(family)에 속하는 바이러스로 유전자형이 매우 다양하다. 본 연구에서는 노로바이러스 국내분리주의 게놈 RNA로부터 3개의 open reading frame (ORF) 모두의 염기서열을 분석하고, 유전학적 계통분석을 통하여 분자생물학적 특성을 분석하였다. 본 연구에 사용된 노로바이러스(Hu/NLV/Gunpo/2006/KO)는 바이러스성 식중독, 장염 증세를 보이는 2세 여아 가검물로부터 분리되었다. 역전사반응과 PCR 증폭을 통해서 바이러스의 게놈 RNA를 3개의 중첩되는 cDNA 단편으로 합성하였으며, 합성된 cDNA를 염기서열 분석에 직접 사용하였다. 시퀀싱 결과 Hu/NLV/Gunpo/2006/KO는 3개의 ORF (ORF1, 5,100 bp; ORF2, 1,647 bp; ORF3, 765 bp)로 구성되어 있음을 알 수 있었다. 35개의 노로바이러스 국외 분리주와 비교한 결과, ORF1은 ORF2 또는 ORF3에 비해서 상대적으로 염기의 변이율이 낮았으며, 특히 ORF2와 ORF3의 C-말단 부위에서 높은 변이율을 관찰하였다. 유전학적 계통도를 분석한 결과, Hu/NLV/Gunpo/2006/KO는 genogroup II 에 속하며, Saitama U1, Gifu'96, Mc37, Vietnam 026과 같은 클러스터를 형성하는 것을 알 수 있었다. 본 연구를 통하여 노로바이러스 Hu/NLV/Gunpo/2006/KO의 3개의 ORF 염기서열을 모두 밝힘으로써, 앞으로 노로바이러스의 검출법 개발과 유전학적 상관관계뿐 아니라, 유전자의 기능 분석과 관련된 기초연구에 중요한 기초자료를 제공할 수 있을 것으로 기대한다.

Keywords

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