杜仲1-羟基-2-甲基-2-(E)-丁烯基-4-二磷酸合酶基因cDNA全长克隆与序列分析  

Cloning and Sequence Analysis of 1-Hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate Synthase Gene cDNA from Eucommia ulmoides

作　　者：刘攀峰[1] 杜红岩[1] 杜兰英[1] 乌云塔娜[2] 黄海燕[1] 

机构地区：[1]中国林业科学研究院经济林研究开发中心,郑州450003 [2]国家林业局经济林育种与栽培重点实验室,长沙410004

出　　处：《植物研究》2012年第4期444-451,共8页Bulletin of Botanical Research

基　　金：国家公益性行业科研专项(201004029)

摘　　要：植物萜类生物合成MEP途径中1-羟基-2-甲基-2-(E)-丁烯基-4-二磷酸合酶(HDS)催化ME-2,4cPP生成HMBPP。以杜仲叶片cDNA为模板,采用反转录RCR及RACE技术分离出HDS基因的cDNA全长克隆。测序及序列分析结果表明该基因全长2 786 bp,基因内部含有完整的开放阅读框,共编码743个氨基酸,推导的蛋白质分子量为82.25 kD,理论等电点为5.89,编码序列含有2个保守的结构域PSN和PSI以及3个绝对保守的半胱氨酸位点。系统进化树分析表明EuHDS蛋白与葡萄HDS蛋白的进化距离最为接近(0.049),其次为番茄(0.052)和橡胶(0.052)。1-Hydroxy-2-methyl-2-（E） -butenyl-4-diphosphate synthase （HDS） catalyses 2C-methyl-D-erythritol- 2,4-cyclodiphosphate into 1-hydroxy-2-methyl-2-（ E）-butenyl-4-diphosphate in the penultimate step of 2C-meth- yl-D-erythritol 4-phosphate （MEP） pathway, which is an alternative plant terpenoids biosynthetic route that has been recently discovered. Homologous HDS eDNA （EuHDS） was isolated from leaves of Eucommia ulrnoides by the methods of reverse transcription polymerase chain reaction （RT-PCR） and rapid amplification of eDNA ends （RACE） technique. The full-length eDNA of EuHDS was 2 786 bp and encoded 743 amino acids with a predicted molecular mass of 82.25 kD, and a theoretical isoelectric point of 5.89. Two conserved motifs and three absolutely conserved cysteines （residues 644, 647 and 678 ） were discovered in the deduced coding sequence. Phylogenetic analysis revealed that EuDXR was more similar to Vitis vinifera （ evolutionary distance, 0.049 ） than that of other species, followed by Solanum lycopersicum （0. 052） and Hevea brasiliensis （0. 052）.

关 键 词：杜仲 HDS 基因 序列分析 

分 类 号：S567.1[农业科学—中草药栽培]