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首页> 外文期刊>DNA Sequence: The Journal of DNA Sequencing and Mapping >Cloning and expression analysis of a vacuolar Na+/H+ antiporter gene from Alfalfa
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Cloning and expression analysis of a vacuolar Na+/H+ antiporter gene from Alfalfa

机译:苜蓿液泡状Na + / H +反转运蛋白基因的克隆和表达分析

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A full-length cDNA, named MsNHX1, encoding a vacuolar Na+/H+ antiporter was cloned from alfalfa (Medicago sativa), using degenerate primers, followed by 3' and 5' RACE. The cDNA sequence was 2225 bp long and included an open reading frame encoding a deduced protein of 541-amino-acid polypeptide. The deduced amino acid sequence showed high similarity (more than 73%) to those of the previously cloned Na+/H+ antiporters; form Arabidopsis thaliana, Qryza sativa, Atriplex gemlinin, Beta vulgaris and Hordeum vulgare. Southern hybridization showed that MsNHX1 has only one copy in alfalfa genome. Semiquantitative RT-PCR analysis indicated that the mRNA level of MsNHX1 was moderate without stress and markedly up-regulated after treatment by NaCl and ABA (abscisic acid). Those results suggest that the MsNHX1 product play an important role in salt tolerance of the alfalfa, and its transcript expression is possibly partially regulated through ABA-dependent signaling pathway.
机译:使用简并引物,3'和5'RACE,从苜蓿(苜蓿)中克隆了一个编码液泡状Na + / H +反转运蛋白的全长cDNA,称为MsNHX1。 cDNA序列长2225 bp,包括一个开放阅读框,编码一个541个氨基酸的多肽推导蛋白。推导的氨基酸序列与先前克隆的Na + / H +反转运蛋白具有很高的相似性(超过73%)。形成拟南芥,紫花苜蓿,滨藜属吉林宁,甜菜和大麦。 Southern杂交表明,MsNHX1在苜蓿基因组中只有一个拷贝。半定量RT-PCR分析表明,经NaCl和ABA(脱落酸)处理后,MsNHX1的mRNA水平为中等水平,无压力,且明显上调。这些结果表明,MsNHX1产物在苜蓿的耐盐性中起重要作用,其转录本表达可能通过ABA依赖性信号传导通路部分调节。

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