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新城疫病毒广西分离株F基因的克隆和序列分析
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唐小飞，谢芝勋，刘加波，庞耀珊，邓显文
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（广西兽医研究所，广西 南宁 530001）
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摘要：
根据GenBank中登录的新城疫病毒（NDV）F基因序列设计了2对引物，对从广西分离的10株NDV毒株的F基因进行了分段扩增和序列测定，其基因序列全长均为1 662 bp，编码553个氨基酸，均有6个潜在的糖基化位点。将其F基因的核苷酸序列及推导的氨基酸序列与已发表的10株NDV参考株的F基因序列进行了比较。结果表明，核苷酸序列的同源性为82.6%～ 98.1%，氨基酸的同源性为87.7%～98.7%。这10株NDV广西分离株在裂解位点的氨基酸序列（112 R-R-Q-K/R-R-F 117 ）与NDV强毒株特征相符合。系统发育树、酶切位点分析和基因分型结果表明，10株NDV广西分离株中GX1/00、GX2/00、GX4/00、GX6/02、GX7/02、GX9/03、GX10/03和GX11/03为基因Ⅶd亚型，GX3/00和GX5/00为基因Ⅲ型。 |
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关键词：
新城疫病毒；F基因；克隆；序列分析 |
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中图分类号：S 852.659.5:Q 785 文献标识码：
A 文章编号：1000-6419(2005)05-0333-08 |
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Cloning and sequencing of fusion protein gene of Newcastle disease virus isolates in Guangxi |
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TANG Xiao-fei，XIE Zhi-xun，LIU Jia-bo，PANG Yao-shan，DENG Xian-wen |
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（Guangxi Veterinary Research Institute，Nanning 530001，China） |
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Abstract:Two pairs of specific primers were designed and synthesized according to fusion （F） protein gene sequences of Newcastle disease virus （NDV） from GenBank. F gene of ten NDV isolates from the chicken farms in Guangxi Region，where outbreak of Newcastle disease had recently occurred，were amplified. All complete nucleotide sequences of the F gene are 1，662 bp in length，which encodes 553 amino acids and contains 6 glycosylation sites. Respective comparison of the ten NDV isolates F gene complete sequences and the deduced amino acid sequences with ten other published NDV F gene complete sequences and the deduced amino acid sequences showed that the homologies of the nucleotide are 82.6%-98.1% and the homologies of the deduced amino acid sequences are 87.7%-98.7%. Their amino acid sequences of the cleavage site region residues（112 R-R-Q-K/R-R-F 117 ） matched to the characteristics of virulent strains. The results of phylogenetic tree，restriction site analysis and genotype characterization showed that eight isolates GX1/00，GX2/00，GX4/00，GX6/02，GX7/02，GX9/03，GX10/03 and GX11/03 belongs to novel subgenotype Ⅶd，and the other two isolates GX3/00 and GX5/00 belongs to the genotype Ⅲ. |
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Key words: Newcastle disease virus；fusion protein gene；cloning；sequencing |
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