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温州蜜柑线粒体二羧酸/三羧酸转运蛋白基因在响应铝胁迫时的分子克隆与特性分析

Molecular cloning and characterization of a mitochondrial dicarboxylate/tricarboxylate transporter gene in Citrus junos response to aluminum stress.

作者信息

Deng Wei, Luo Keming, Li Zhengguo, Yang Yingwu

机构信息

Key Laboratory of Functional Gene and Regulation Technologies under Chongqing Municipal Education Commission, Bioengineering College, Chongqing University, Genetic Engineering Research Center, Chongqing, People's Republic of China.

出版信息

Mitochondrial DNA. 2008 Aug;19(4):376-84.

Abstract

A mitochondrial dicarboxylate/tricarboxylate carrier gene, CjDTC, was isolated from Citrus junos by the rapid amplification of cDNA ends and the Y-shaped adaptor-dependent extension methods. It consisted of a 472-base pair (bp) upstream regulatory region and an 897-bp open reading frame encoding a protein of 299 amino acids. Homologous analysis revealed that CjDTC protein might be a plant dicarboxylate/tricarboxylate carrier protein involved in the mitochondria dicarboxylate/tricarboxylate transport. The putative light responsiveness and salicylic acid responsiveness regulatory elements were identified in the upstream regulatory region of CjDTC. Southern blot analysis demonstrated the presence of a single CjDTC gene located on the genome of citrus. As shown by northern hybridization, CjDTC was expressed in all plant tissues examined and the highest transcript level was observed in roots with significantly lower transcript amounts in leaves and stems. Moreover, real-time polymerase chain reaction analysis demonstrated that CjDTC expression was induced by aluminum treatment, suggesting that CjDTC protein might be involved in the excretion of organic acids and rhizotoxic aluminum tolerance.

摘要

通过cDNA末端快速扩增法和Y形接头依赖延伸法,从日本柑橘中分离出一个线粒体二羧酸/三羧酸载体基因CjDTC。它由一个472个碱基对(bp)的上游调控区和一个897 bp的开放阅读框组成,该开放阅读框编码一个含299个氨基酸的蛋白质。同源性分析表明,CjDTC蛋白可能是一种参与线粒体二羧酸/三羧酸转运的植物二羧酸/三羧酸载体蛋白。在CjDTC的上游调控区鉴定出了假定的光响应和水杨酸响应调控元件。Southern杂交分析表明,柑橘基因组中存在一个单一的CjDTC基因。Northern杂交显示,CjDTC在所检测的所有植物组织中均有表达,在根中观察到最高转录水平,在叶和茎中的转录量显著较低。此外,实时聚合酶链反应分析表明,铝处理可诱导CjDTC表达,这表明CjDTC蛋白可能参与有机酸的分泌和对根际毒性铝的耐受性。

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