基因VIb亚型半番鸭源新城疫病毒基因组测序及分析
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广东省兽医公共卫生公共实验室开放基金(GSKJ090201)


Sequencing and Analysis Genome of a VIb Subgenotype of Newcastle Disease Virus Isolated from Semi-Muscovy Duck
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    摘要:

    根据GenBank上发布的新城疫病毒基因组序列,设计了9对引物,对从福建福州地区发病半番鸭场分离的新城疫病毒FJ-SMD-03株进行了基因组cDNA扩增测序和分析。结果表明:FJ-SMD-03的基因组序列由15 192 nt组成,在NP基因非编码区比Lasota株多了6个核苷酸ACACTC;基因组由6个ORF组成,即3′-NP-P-M-F-HN-L-5′序列。BLAST结果显示:FJ-SMD-03基因组核苷酸与1.3、carrier-pigeon/Guangdong/2008、pigeon/Italy/1166/00、AV324/96 和dove/Italy/2736/00相似性最高,达94.9%~98.5%;与疫苗株Lasota、Mukteswar和B1的相似性最低,为80.7%~83.3%;与我国标准强毒株F48E8及2002年从福建番鸭中分离到的基因VIId亚型强毒株Muscovy duck/China(Fujian)/FP1/02相似性较低,分别为82.5%和87.1%。F蛋白裂解位点的氨基酸序列为112R-R-Q-K-R-F117,具有强毒株序列特征,进化树分析发现,FJ-SMD-03属于Class Ⅱ类基因VIb亚型,证明鸭群中有VIb亚型NDV强毒株;FJ-SMD-03株NDV与1.3、carrier-pigeon/Guangdong/2008和pigeon/Italy/1166/00亲缘关系较近,与carrier-pigeon/Guangdong/2008形成一个小的进化分支,推测FJ-SMD-03株与carrier-pigeon/Guangdong/2008和pigeon/Italy/1166/00株来源于共同的祖代毒株,可能与鸽携带的病毒有关。

    Abstract:

    Based on the genome sequence of newcastle disease virus(NDV) published in GenBank,nine pairs of gene-specific primers were designed to amplify cDNA of FJ-SMD-03 strain isolated from Semi-Muscovy duck in Fuzhou, Fujian ,then sequecing and analysis were performed. The results showed that FJ-SMD-03 was 15 192 nucleotides in length which consisted of six ORFs in the order of 3′leader-NP-P-M-F-HN-L-5′trailer and it had six nucleotides longer than Lasota strain in non-coding region of NP gene. The genome shared the highest similarity of the nucleotide with 1.3,carrier-pigeon/Guangdong/2008,pigeon/Italy/1166/00,AV324/96 and dove/Italy/2736/00 (94.9%-98.5%),and shared the lowest similarity of the nucleotide with vaccine strain Lasota,Mukteswar and B1(80.7%-83.3%), whereas it shared lower similarity of the nucleotide with strain F48E8 and Muscovy duck/China(Fujian)/FP1/02,Subgenotype VIId, velogenic, isolated from Muscovy duck in Fujian during 2002 for 82.5% and 87.1% respectively. The motif of cleavage site region of F gene was 112R-R-Q-K-R-F117, which was the sequence signature of virulent strain. Phylogenetic analysis revealed that FJ-SMD-03 belonged to subgenotype VIb, Class Ⅱ, which was the first report of the subgenotype VIb virulent NDV strain isolated from Semi-Muscovy duck. FJ-SMD-03 was closely associated with 1.3,carrier-pigeon/Guangdong/2008 and pigeon/Italy/1166/00 and formed a secondary branch in the phylogenetic tree with carrier-pigeon/Guangdong/2008, which suggested that FJ-SMD-03, carrier-pigeon/Guangdong/2008 and pigeon/Italy/1166/00 came from the same origin, which might be due to virus carried by pigeons.

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黄秋芳,孙敏华,陈少莺,吕殿红,董嘉文,尚毅,吴玄光,胡奇林.基因VIb亚型半番鸭源新城疫病毒基因组测序及分析[J].华南农业大学学报,2012,33(1):106-112

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  • 收稿日期:2011-06-02
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  • 在线发布日期: 2012-04-13