茶树金属耐受蛋白基因CsMTP11的克隆及功能分析

发布时间：2018-02-21 21:51

本文关键词： 茶树重金属耐受蛋白基因克隆锰　出处：《作物学报》2017年05期 　论文类型：期刊论文

【摘要】：金属耐受蛋白MTP(metal tolerance protein)是阳离子转运蛋白(CDF)家族的重要成员,在植物重金属转运过程中发挥重要调控作用。本研究以中茶108茶树为试验材料,通过RT-PCR和RACE方法克隆到茶树重金属耐受蛋白基因CsMTP11(Gen Bank登录号为KX450265),其全长c DNA为1197 bp,编码398个氨基酸残基,其编码蛋白分子量为44.85 k D,等电点为5.34。在线软件分析表明,CsMTP11蛋白具有5个跨膜结构域,且含有CDF家族的其他保守结构域。系统进化树分析结果表明,茶树CsMTP11与葡萄VvMTP11进化同源性最近,其氨基酸序列相似度高达90%。基因表达模式分析表明,CsMTP11基因在茶树老叶中的表达量最高,根中的表达量最低,另外,CsMTP11基因受重金属Mn和Co离子胁迫诱导表达。CsMTP11-YFP融合蛋白在拟南芥原生质体共定位试验表明,CsMTP11-YFP融合蛋白定位于质膜。CsMTP11在酿酒酵母及其突变株中的异源表达可以提高其对重金属Mn和Co离子的耐受性。综上所述,茶树CsMTP11属于Mn-CDF亚家族,可能参与茶树对重金属锰和钴的转运过程。
[Abstract]:Metal tolerance protein (MTP(metal tolerance) is an important member of cationic transporter protein family, which plays an important role in heavy metal transport in plants. In this study, Zhongcha 108 tea plant was used as experimental material. By means of RT-PCR and RACE, the CsMTP11(Gen Bank gene of tea tree heavy metal tolerance protein was cloned into KX450265, and its full-length c DNA was 1197 BP, encoding 398 amino acid residues. The molecular weight of the encoded protein is 44.85 KD and the isoelectric point is 5.34. The on-line software analysis shows that the CsMTP11 protein has five transmembrane domains and other conserved domains of the CDF family. Phylogenetic tree analysis shows that CsMTP11 protein has five transmembrane domains. The similarity of amino acid sequence of tea plant CsMTP11 and grape VvMTP11 is as high as 90%. The expression pattern of CsMTP11 gene is the highest in the old leaves of tea plant and the lowest in the root. In addition, the co-localization of CsMTP11-YFP fusion protein induced by heavy metal mn and Co ions stress in Arabidopsis thaliana protoplasts showed that CsMTP11-YFP fusion protein was located in plasma membrane. CsMTP11 could be expressed in Saccharomyces cerevisiae and its mutant. High tolerance to heavy metal mn and Co ions. Tea CsMTP11 belongs to the Mn-CDF subfamily and may be involved in the transport of heavy metals mn and Co in tea plants.
【作者单位】：西南大学食品科学学院;
【基金】：国家自然科学基金项目(31400583) 重庆市基础与前沿研究计划一般项目(cstc2014jcyjA80011) 中央高校基本业务费专项(XDJK2014C070)资助~~
【分类号】：S571.1

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