Korean Journal of Veterinary Service (한국동물위생학회지)

The Korean Society of Veterinary Service (한국동물위생학회)
권호 표지

Molecular cloning and nucleotide sequence of the gene encoding hemagglutinin-neuraminidase(HN) of Newcastle disease virus isolated from a diseased pheasant in Korea

국내 사육 꿩에서 분리된 뉴켓슬병 바이러스의 hemagglutinin-neuraminidase(HN) 유전자의 클론닝과 염기서열 분석

  • 장경수 (전라북도축산기술연구소) ;
  • 곽길한 (전라북도축산기술연구소) ;
  • 장승익 (대전광역시보건환경연구원) ;
  • 김지영 (전북대학교 수의과대학 및 생체안전성연구소) ;
  • 김태용 (전북대학교 수의과대학 및 생체안전성연구소) ;
  • 송영환 (전북대학교 수의과대학 및 생체안전성연구소) ;
  • 송희종 (전북대학교 수의과대학 및 생체안전성연구소) ;
  • 전무형 (충남대학교 수의과대학)
  • Published : 2002.09.01
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Abstract

The gene encoding the HN protein from the CBP-1 strain, a heat stable Newcastle disease virus (NDV) isolated from diseased pheasants in Korea, was characterized by reverse transcriptase- polymerase chain reaction(RT-PCR) and the nucleotide and amino acid sequences were analyzed following cloning of the HN gene. In all of the NDV strains studied, a 1.75 kb size cDNA fragment for the HN gene was generated by RT-PCR and smaller specific band sizes harboring the internal portions of the HN gene were also detected by using four pairs of primers. The RT-PCR was sensitive enough to detect viral transcripts when the virus titer was above 25 hemagglutination units. The amplified 1.75 kb cDNA was cloned into a BamHI site of the pVL1393 Baculo transfer vector. The nucleotide sequences of the 1,758 bp HN gene from the CBP-1 strain were determined by the dye terminator cyclic sequencing method. The gene sequences were compared among the strains of CBP-1, Texas GB, Beaudette C, LaSota, B1 and Ulster. The homology of the CBP-1 HN gene to other HN variants was 97.8% to Texas GB, 98.4% to Beaudette C, 95.4% to LaSota, 95.6% to B1 and 90.2% to Ulster. As the deduced 577 amino acid sequences were compared among the strains, the homology for CBP-1 HN appeared to be 96.7% to Texas GB, 97.9% to Beaudette C, 95.5% to LaSota, 95.5% to B1 and 92.7% to Ulster. It was evident that the amino acid sequences included 5 sites for N-asparagine linked glycosylation and 12 cysteine residues. The three conserved leucine residues within the predicted transmembrane domain of the HN protein are amino acid 30, 37 and 44. The three antigenic sites on the HN protein of NDV are amino acids 347(Glu), 481(Asn) and 495(Glu). These data indicate that the genotype of the CBP-1 strain is more closely associated with the strains of Texas GB and Beaudette C than it is for the LaSota, B1 and Ulster strains.

Keywords

References

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