研究论文

小麦TaCAT新基因克隆及分子生物学和生化特性分析

  • 佘茂云;雷 昊;殷桂香;冯 晨;别晓敏;杜丽璞;徐惠君;叶兴国
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  • 中国农业科学院作物科学研究所;国家农作物基因资源与基因改良重大科学工程;农业部作物遗传育种重点开放实验室,北京 100081

收稿日期: 2011-02-23

  修回日期: 2011-03-29

  网络出版日期: 2011-04-22

Clone of TaCAT Genes in Wheat (Triticum aestivum L.) and Its Biochemical and Molecular Biological Characterization

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Received date: 2011-02-23

  Revised date: 2011-03-29

  Online published: 2011-04-22

摘要

小麦组织培养再生潜力与抗氧化胁迫能力具有相关性。为解析小麦TaCATs基因家族的分子生物学及生化特性并为后续试验验证奠定理论基础,利用e-PCR方法进行小麦过氧化氢酶(CAT)新基因克隆,结合in silico技术对小麦TaCATs基因家族的生化特性进行分析和预测。氨基酸序列同源性比对结果表明,克隆的小麦TaCATs基因家族新成员与水稻的CatA和玉米的Cat3具有较高的相似性,分别达89%及81.1%,命名为TaCAT3,基因组DNA和cDNA长度分别为1986和1482bp,编码494个氨基酸的蛋白。亚细胞定位结果表明,TaCAT3可能定位在线粒体中,且所有的功能活性位点在小麦TaCATs家族中具有一致的保守性。系统发生树构建结果表明,小麦TaCATs能形成3个独立的分支。在蛋白质编码序列同源比对的基础上,利用SwissModel的Swiss-PdbViewer 3.7软件包对TaCATs高级结构进行同源模拟,发现所构建的模型能很好地反映TaCATs的高级结构。

本文引用格式

佘茂云;雷 昊;殷桂香;冯 晨;别晓敏;杜丽璞;徐惠君;叶兴国 . 小麦TaCAT新基因克隆及分子生物学和生化特性分析[J]. 科技导报, 2011 , 28(11-12) : 23 -31 . DOI: 10.3981/j.issn.1000-7857.2011.12.002

Abstract

There is a close relationship between in vitro regeneration potential of wheat (Triticum aestivum L.) tissues and their resistant ability to oxidative stress. In order to dissect and provide theoretical evidences for subsequent experimental molecular functional characterization of TaCATs, e-PCR method and in silico techniques are employed for the isolation of the new member of wheat TaCAT family and the prediction on biochemical characters of TaCATs, respectively. TaCAT3, as a new member in wheat CAT gene family, is encoded a deduced 494-amino acid protein with the 1986bp and 1482bp of genomic DNA and cDNA, respectively, it shares 89% homology with CatA (Oryza sativa L.) and 81.1% identity with Cat3 (Zea mays L.), and is possibly targeted in the mitochondria according to the subcellular localization prediction. Furthermore, all the functional sites in TaCATs are highly conserved. Phylogenetic tree construction shows that the wheat TaCATs are able to form three separate branches with one isoform each. Swiss-PdbViewer 3.7 software package is employed for homology modeling of TaCATs after an appropriate template is acquired in Swiss Model online. The reconstructed model could well reflect the TaCAT senior structure.
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