烟草果胶甲基酯酶(PME)基因的克隆及功能分析(英)
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国家杰出人才科学基金项目(30125004),国家重点基础研究发展规划项目(973)(G200000016201)和国家211工程资助项目.


Cloning and Functional Analysis of Tobacco Pectin Methylesterase
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This work was supported by grants from National Science Fund for Distinguished Young Scholars (30125004), The Special Funds for Major State Basic Research of China (G200000016201 ) and National 211 Project.

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    摘要:

    为了研究果胶甲基酯酶(pectin methyl-esterase,PME)(EC 3.1.1.11)与植物病毒运动蛋白之间的相互作用,应用RT-PCR方法从烟草(Nicotiana benthamiana)中克隆了PME基因,并测定了全序列(GenBank登录号AY238968).序列分析显示该基因由两个保守的结构域组成(PMEI和pectinesterase). DNA印迹结果表明,该基因在基因组中存在多个拷贝,蛋白质印迹表明,植物总蛋白中存在两种形式的PME蛋白,但RNA印迹结果显示,在烟草细胞中只检测到全长的PME转录产物.酵母双杂交结果表明,PME与水稻矮缩病毒Pns11(具有非特异的核酸结合活性)之间存在相互作用,而没有检测到PME与已知的运动蛋白Pns6之间的相互作用,推测Pns11蛋白可能参与了水稻矮缩病毒粒子的运动.

    Abstract:

    In order to study the interactions between plant virus movement protein and pectin methylesterase (PME), PME gene from tobacco genome was cloned using RT-PCR method and the sequence was determined (GenBank accession No.AY238968). Sequence analysis showed that there were two conserved domains in PME protein: PMEI and pectinesterase. Multiple copies of PME gene were detected in tobacco genome through Southern blot analysis. Western blot result indicated that two types of PME protein existed in tobacco. But Northern hybridization detected only a full length transcript of PME, which further identified that there was post-translational process. Yeast two-hybrid result demonstrated that PME could not interact with the identified movement protein of rice dwarf virus (RDV), Pns6, but does interact with Pns11, a nucleic acid binding protein of RDV. It implies Pns11 may participate in the movement of RDV.

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李春波,高锋,钟永旺,魏春红,李毅.烟草果胶甲基酯酶(PME)基因的克隆及功能分析(英)[J].生物化学与生物物理进展,2004,31(7):643-649

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  • 收稿日期:2003-12-23
  • 最后修改日期:2004-01-10
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