张琳, 王继涛, 张大为, 张岗, 郭顺星. 珍稀濒危药用铁皮石斛HMGR基因的克隆和特征分析J. 药学学报, 2014,49(3): 411-418.
引用本文: 张琳, 王继涛, 张大为, 张岗, 郭顺星. 珍稀濒危药用铁皮石斛HMGR基因的克隆和特征分析J. 药学学报, 2014,49(3): 411-418.
ZHANG Lin, WANG Ji-tao, ZHANG Da-wei, ZHANG Gang, GUO Shun-xing. Molecular characterization of a HMG-CoA reductase gene from a rare and endangered medicinal plant, Dendrobium officinaleJ. Acta Pharmaceutica Sinica, 2014,49(3): 411-418.
Citation: ZHANG Lin, WANG Ji-tao, ZHANG Da-wei, ZHANG Gang, GUO Shun-xing. Molecular characterization of a HMG-CoA reductase gene from a rare and endangered medicinal plant, Dendrobium officinaleJ. Acta Pharmaceutica Sinica, 2014,49(3): 411-418.

珍稀濒危药用铁皮石斛HMGR基因的克隆和特征分析

Molecular characterization of a HMG-CoA reductase gene from a rare and endangered medicinal plant, Dendrobium officinale

  • 摘要: 3-羟基-3-甲基戊二酸单酰辅酶A还原酶(3-hydroxy-3-methylglutaryl coenzyme-A reductase,HMGR)是甲羟戊酸途径中的第一个限速酶,在植物细胞萜类物质合成代谢过程中发挥重要作用。但是,HMGR基因在珍稀濒危兰科药用植物铁皮石斛类萜生物合成途径中的作用尚不清楚。本研究利用RT-PCR和RACE技术,首次从铁皮石斛中克隆到一个HMGR基因,命名为DoHMGR1(GenBank注册号JX272632)。DoHMGR1基因cDNA全长2 071 bp,编码一条由562个氨基酸组成的多肽,分子量59.73 kD,等电点6.18。DoHMGR1蛋白包含HMGR蛋白家族的4个保守结构域(63~561、147~551、268~383、124~541)和两个跨膜基序(42~64、85~107)。DoHMGR1基因与多种植物HMGR基因相似性很高(67%~89%),与万代兰、水稻、玉米等单子叶植物亲缘关系较近。DoHMGR1基因具有组织表达特异性,其转录本在石斛根和茎中表达量较高,为叶中的2.13和1.98倍。DoHMGR1基因的分子鉴定为进一步解析该基因在铁皮石斛萜类物质合成代谢途径中的分子调控作用奠定基础。

     

    Abstract: The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) catalyzes the conversion of HMG-CoA to mevalonate in mavalonic acid pathway, which is the first committed step for isoprenoid biosynthesis in plants. However, it still remains unclear whether HGMR gene plays a role in the isoprenoid biosynthesis in Dendrobium officinale, an endangered epiphytic orchid species. In the present study, a HMGR encoding gene, designed as DoHMGR1 (GenBank accession JX272632), was identified from D. officinale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods, for the first time. The full length cDNA of DoHMGR1 was 2 071 bp in length and encoded a 562-aa protein with a molecular weight of 59.73 kD and an isoelectric point (pI) of 6.18. The deduced DoHMGR1 protein, like other HMGR proteins, constituted four conserved domains (63-561, 147-551, 268-383 and 124-541) and two transmembrane motifs (42-64 and 85-107). Multiple sequence alignment and phylogenetic analyses demonstrated that DoHMGR1 had high identity (67%-89%) to a number of HMGR genes from various plants and was closely related to Vanda hybrid cultivar, rice and maize monocots. Real time quantitative PCR (qPCR) analysis revealed that DoHMGR1 was expressed in the three included organs. The transcripts were the most abundant in the roots with 2.13 fold over that in the leaves, followed by that in the stems with 1.98 fold. Molecular characterization of DoHMGR1 will be useful for further functional elucidation of the gene involving in isoprenoid biosynthesis pathway in D. officinale.

     

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