长叶红砂液泡膜Na+/H+逆向转运蛋白基因的克隆及表达特性
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国家自然科学基金(31360063);内蒙古自治区高等学校创新研究团队发展计划(NMGIRT1401)


Isolation and Expression of Vacuolar Membrane Na+/H+ Antiporter Gene in Reaumuria trigyna
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    摘要:

    为研究长叶红砂(Reaumuria trigyna)离子转运分子机制,利用RT-PCR和RACE技术,克隆到其液泡膜Na+/H+逆向转运蛋白基因(NHX1)的全长cDNA片段,命名为RtNHX1(NCBI序列号为KR919802)。 结果表明:RtNHX1的cDNA片段全长2 622 bp,开放阅读框1 662 bp,5′非编码区509 bp,3′非编码区451 bp,编码553个氨基酸,推测分子量为60.91 kD。该蛋白含有12个跨膜结构域,为疏水蛋白,与其他植物液泡膜Na+/H+逆向转运蛋白NHX1的亲缘关系较近。实时荧光定量PCR对其在NaCl胁迫下的表达检测显示,不同时间和不同浓度NaCl胁迫下,RtNHX1表达量变化均呈先升高后降低趋势,在100 mmol/L NaCl胁迫6 h和200 mmol/L NaCl胁迫后达到最高,表达量分别超过或约是对照的3倍,一定程度反应出RtNHX1参与长叶红砂的盐胁迫应答,是该植物离子转运体系的重要元件。

    Abstract:

    To investigate the molecular mechanism of the ion transport in Reaumuria trigyna,we cloned the full length cDNA of vacuolar membrane Na+/H+ antiporter gene(RtNHX1) with reverse transcription PCR and rapid amplification of cDNA ends technologies.The cDNA is 2 622 bp long,which contained a 1 662 bp open reading frame,a 509 bp 5′-UTR,and a 451 bp 3′-UTR.The cDNA encoded a protein with 553 amino acids and molecular weight of 60.91 kD.The protein was predicted to be composed of 12 transmembrane domains,was characterized as a hydrophobic protein,and showed high homology with the vacuolar membrane Na+/H+ antiporter(NHX1) in other plant species.Quantitative real-time analysis showed that the expression levels of RtNHX1 under different salt concentrations and time gradients,were up-regulated initially and then down-regulated.The expression of RtNHX1 under the stress of 100 mmol/L NaCl for 6 hours and 200 mmol/L NaCl were reached the highest,which were over or nearly 3-fold of the control,indicating that RtNHX1 participated in the salt stress response of R.trigyna and may be an important element that plays a role in the ion transporting system in this plant.

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董禄禄,秦晓春,党振华.长叶红砂液泡膜Na+/H+逆向转运蛋白基因的克隆及表达特性[J].西北植物学报,2015,35(11):2164-2170

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  • 在线发布日期: 2015-12-07
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