School of Chemical Engineering, China University of Mining and Technology, Xuzhou, People's Republic of China.
Mol Biol Rep. 2010 Mar;37(3):1559-67. doi: 10.1007/s11033-009-9558-7. Epub 2009 May 13.
A new full-length cDNA encoding 3-hydroxy-3-methylglutoryl-Coenzyme A reductase (HMGR; EC1.1.1.34), which catalyzes the first committed step of isoprenoids biosynthesis in MVA pathway, was isolated from young leaves of Euphorbia Pekinensis Rupr. by rapid amplification of cDNA ends (RACE) for the first time. The full-length cDNA of HMGR (designated as EpHMGR, GenBank Accession NO. EF062569) was 2,200 bp containing a 1,752 bp ORF encoding 583 amino acids. Bioinformatic analyzes revealed that the deduced EpHMGR had extensive homology with other plant HMGRs and contained two transmembrane domains and a catalytic domain. The predicted 3-D model of EpHMGR had a typical spatial structure of HMGRs. Southern blot analysis indicated that at most two copies of EpHMGR gene existed in E. Pekinensis genome. Tissue expression analysis revealed that EpHMGR expressed strongly in roots, weakly in stems and leaves. The functional colour complementation assay indicated that EpHMGR could accelerate the biosynthesis of carotenoids in the Escherichia coli transformant, demonstrating that EpHMGR plays an influential role in isoprenoid biosynthesis.
我们首次通过快速扩增 cDNA 末端（RACE）技术，从大戟属植物Euphorbia Pekinensis Rupr. 的幼叶中分离到编码 3-羟基-3-甲基戊二酰辅酶 A 还原酶（HMGR；EC1.1.1.34）的全长 cDNA，该酶催化甲羟戊酸途径中异戊烯基生物合成的第一步。HMGR 的全长 cDNA（命名为 EpHMGR，GenBank 登录号 EF062569）长 2200bp，包含一个编码 583 个氨基酸的 1752bpORF。生物信息学分析表明，推导的 EpHMGR 与其他植物 HMGR 具有广泛的同源性，并且包含两个跨膜结构域和一个催化结构域。EpHMGR 的预测 3-D 模型具有 HMGR 典型的空间结构。Southern blot 分析表明，EpHMGR 基因在大戟属基因组中最多存在两个拷贝。组织表达分析表明，EpHMGR 在根中表达强烈，在茎和叶中表达较弱。功能颜色互补测定表明，EpHMGR 可以加速大肠杆菌转化体中类胡萝卜素的生物合成，表明 EpHMGR 在异戊烯基生物合成中发挥重要作用。
