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首页> 外文期刊>Journal of genetics >Molecular cloning, characterization and expression analysis of the gene encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase from Aquilaria sinensis (Lour.) Gilg
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Molecular cloning, characterization and expression analysis of the gene encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase from Aquilaria sinensis (Lour.) Gilg

机译：中国沉香（Aquilaria sinensis（Lour。）Gilg）1-脱氧-D-木酮糖5-磷酸还原异构酶编码基因的分子克隆，表征和表达分析

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摘要

The major constituents of agarwood oils are sesquiterpenes that are obtained from isoprenoid precursors through the plastidial methylerythritol phosphate (MEP) pathway and the cytosolic mevalonate pathway. In this study, a novel full-length cDNA of 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR), which was the second key enzyme in the plastid MEP pathway of sesquiterpenes biosynthesis was isolated from the stem of Aquilaria sinensis (Lour.) Gilg by the methods of reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) technique for the first time, and named as AsDXR. The full-length cDNA of AsDXR was 1768 bp, containing a 1437 bp open reading frame (ORF) encoding a polypeptide of 478 amino acids with a molecular weight of 51.859 kD and the theoretical isoelectric point of 6.29. Comparative and bioinformatic analysis of the deduced AsDXR protein showed extensive homology with DXRs from other plant species, especially Theobroma cacao and Gossypium barbadense, and contained a conserved transit peptide for plastids, and extended pro-rich region and a highly conserved NADPH-binding motif owned by all plant DXRs. Southern blot analysis indicated that AsDXR belonged to a small gene family. Tissue expression pattern analysis revealed that AsDXR expressed strongly in root and stem, but weakly in leaf. Additionally, AsDXR expression was found to be activated by exogenous elicitor of MeJA (methyl jasmonate). The contents of three sesquiterpenes (alpha-guaiene, alpha-humulene and delta-guaiene) were significantly induced by MeJA. This study enables us to further elucidate the role of AsDXR in the biosynthesis of agarwood sesquiterpenes in A. sinensis at the molecular level.
机译：沉香油的主要成分是倍半萜烯，其是通过质体甲基赤藓糖醇磷酸酯（MEP）途径和胞质甲羟戊酸途径从类异戊二烯前体获得的。在这项研究中，从沉香（Aquilaria sinensis）的茎中分离了一种新的全长cDNA 1-脱氧-D-木酮糖5-磷酸还原异构酶（DXR），这是倍半萜生物合成质体MEP途径中的第二个关键酶。）Gilg首次采用逆转录聚合酶链反应（RT-PCR）和cDNA末端快速扩增（RACE）技术，并命名为AsDXR。 AsDXR的全长cDNA为1768 bp，包含一个1437 bp的开放阅读框（ORF），编码478个氨基酸的多肽，分子量为51.859 kD，理论等电点为6.29。推导的AsDXR蛋白的比较和生物信息学分析表明，它与其他植物物种（尤其是可可可氏菌和巴布斯棉）的DXR具有广泛的同源性，并包含用于质体的保守转运肽，并具有丰富的前富集区域和高度保守的NADPH结合基序所有工厂的DXR。 Southern印迹分析表明AsDXR属于一个小基因家族。组织表达模式分析表明，AsDXR在根和茎中表达强，而在叶中表达弱。此外，发现AsDXR表达被MeJA（茉莉酸甲酯）的外源激发子激活。 MeJA显着诱导了三种倍半萜的含量（α-番石榴烯，α-humulene和δ-番石榴烯）。这项研究使我们能够在分子水平上进一步阐明AsDXR在沉香木沉香倍半萜生物合成中的作用。

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