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鸭瘟病毒TK基因的克隆及其分子特性分析
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葛菡1，徐超1，程安春1，2*，汪铭书1，2*，贾仁勇2，朱德康1，2，罗启慧2，陈孝跃1
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（1．四川农业大学 动物医学院 禽病防治研究中心，四川 雅安625014；
2．动物疫病与人类健康四川省重点实验室，四川 雅安625014）
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摘要：
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通过测定动物疫病与人类健康四川省重点实验室构建的鸭瘟病毒（duck plaque virus,DPV）DNA基因文库中重组质粒的DNA序列，得到了该病毒胸苷激酶（TK）基因的ORF。采用PCR扩增出了DPV TK基因并将其克隆到pMD18-T载体上，之后对重组质粒进行了PCR和双酶切(BamHⅠ+HindⅢ)鉴定，并利用生物信息学软件Genscan、ProtScale、SignalP2.0、Scansite、TMpred、Prosite、DNAStar以及在线Predictprotein等分析了TK基因的分子特性。结果显示，DPV TK具有疱疹病毒的典型特征，含有ATP结合结构域和核苷酸结合结构域2个功能结构域，且具有与功能相关的磷酸化位点和氨酰化位点；编码的多肽链中亲水区域大于疏水区域，是一种膜外蛋白。DPV TK基因与禽类疱疹病毒（α-疱疹病毒）的进化关系最近。 |
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关键词：
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鸭瘟病毒；胸苷激酶基因；分子特性 |
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中图分类号：S 852.659.1 文献标识码：
A 文章编号：1673-4696(2008)04-0297-06 |
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Cloning of duck plague virus TK gene and analysis
of its molecular characteristics |
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GE Han1,XU Chao1,CHENG An-chun1，2,WANG
Ming-shu1，2,JIA Ren-yong2,
ZHU De-kang1，2,LUO Qi-hui2,CHEN Xiao-yue1 |
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(1.Research Center for Avian Diseases,College of Veterinary Medicine,Sichuan Agricultural University,Ya’an 625014,
China;2.Key Laboratory of Animal Diseases and Human Health of Sichuan Province,Ya’an 625014,China) |
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Abstract: The open reading frame(ORF) of thymidine kinase(TK) gene from duck plague virus (DPV) was obtained by sequencing the recombinant plasmid selected from a DPV DNA library constructed by the Key Laboratory of Animal Diseases and Human Health of Sichuan Province.DPV TK gene was amplified by PCR and cloned into pMD18-T vector.The positive plasmid was selected and identified by PCR.The DPV TK gene was analyzed by bioinformatics tools of Genscan,ProtScale,SignalP2.0,Scansite,TMpred,Prosite,DNAStar and Predictprotein.The results showed that DPV TK gene contained ATP-binding functional domain and nucleotide binding functional domain.The polypeptide had phosphorylation sites,aminoacyl sites,hydrophilic regions and hydrophobic regions.DPV TK was predicted to be located outside of membrane.Phylogenetic studies demonstrated that DPV TK gene was more closely related to TK of poultry herpesvirus(alpha herpesvirus).
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Key words: duck plague virus(DPV);thymidine kinase gene;molecular characteristic |
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