华北农学报 ›› 2014, Vol. 29 ›› Issue (3): 46-51. doi: 10.7668/hbnxb.2014.03.010

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猪沙波病毒CH430株p70基因克隆与生物信息学分析

刘伟1, 王恩丽1,2, 柳纪省1, 杨彬1, 兰喜1   

  1. 1. 中国农业科学院 兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, 草食动物疫病重点开放实验室, 甘肃 兰州 730046;
    2. 甘肃农业大学 动物医学院, 甘肃 兰州 730070
  • 收稿日期:2014-03-02 出版日期:2014-06-28
  • 作者简介:刘伟(1986-),女,河北唐山人,硕士,主要从事动物传染病分子生物学研究。

Cloning and Bioinformatics Analysis of Porcine Sapovirus CH430 Non-structural Protein p70 Gene

LIU Wei1, WANG En-li1,2, LIU Ji-xing1, YANG Bin1, LAN Xi1   

  1. 1. State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Grazing Animal Diseases, Lanzhou Veterinary Research Institute Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;
    2. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China
  • Received:2014-03-02 Published:2014-06-28

摘要: 利用RT-PCR方法从猪沙波病毒CH430株中克隆出非结构蛋白p70基因并测序,结果显示,p70基因全长1 995 nt,编码665个氨基酸,经同源性比对及遗传进化树分析表明,CH430株为基因Ⅲ型沙波病毒毒株。生物信息学分析显示该蛋白理论等电点(pI)为5.96,理论分子质量为72 876.6 u;其序列上共发现25个磷酸化位点,分别为Ser(6)、Thr(13)、Tyr(6),而蛋白的磷酸化与信号传导有关,预测该蛋白为一重要的信号传导分子;无信号肽和跨膜区;该蛋白含有胰蛋白酶样丝氨酸蛋白酶和RdRp功能域;胰蛋白酶样丝氨酸蛋白酶功能域核心结构为1个由7个β折叠形成柱状结构和1个由1个α螺旋和7个β折叠扭曲平行形成1个不完全的桶状结构共同组成,RdRp功能域核心结构为1个具有3个子结构域空间结构,该结构与右手掌在形态上非常相似,而3个子结构域也被认为是手指、手掌和拇指。

关键词: 猪沙波病毒, 非结构蛋白p70, 克隆, 生物信息学分析

Abstract: The non-structural protein p70 gene of PoSaV CH430 strain was cloned by RT-PCR and then sequenced in our study.The results showed that the full length of p70 gene was 1 995 nt encoding a protein of 665 aa.Homology alignment and evolutionary tree showed that PoSaV CH430 strain belong to GⅢ.The bioinformatics analysis demonstrated that the isoelectric point and molecular weight of non-structural protein p70 were 5.96 and 72 876.6 u.The protein had no signal peptide and transmenbrane domain.There were 25 phosphorylation sites including 6 Sers, 13 Thrs and 6 Tyrs.Protein phosphorylation was concerned with signal transduction, so this protein may be a signaling molecule.There were Trypsin-like serine proteases and RdRp functional domain.Main structure of trypsin-like serine proteases functional domain was a column structure and an incomplete β-barrel.Main structure of RdRp functional domain was a fold whose organisation has been likened to the shape of a right hand with three subdomains termed fingers, palm and thumb.

Key words: Porcine sapovirus, Non-structural protein p70, Clone, Bioinformatics analysis

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引用本文

刘伟, 王恩丽, 柳纪省, 杨彬, 兰喜. 猪沙波病毒CH430株p70基因克隆与生物信息学分析[J]. 华北农学报, 2014, 29(3): 46-51. doi: 10.7668/hbnxb.2014.03.010.

LIU Wei, WANG En-li, LIU Ji-xing, YANG Bin, LAN Xi. Cloning and Bioinformatics Analysis of Porcine Sapovirus CH430 Non-structural Protein p70 Gene[J]. journal1, 2014, 29(3): 46-51. doi: 10.7668/hbnxb.2014.03.010.