Molecular cloning,characterization and function analysis of the gene encoding HMG-CoA reductase from <Emphasis Type="Italic">Euphorbia Pekinensis</Emphasis> Rupr |
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Authors: | Xiaoying Cao Zhimin Zong Xiuyun Ju Yong Sun Chuanchao Dai Qun Liu Jihong Jiang |
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Institution: | (1) School of Chemical Engineering, China University of Mining and Technology, 221008 Xuzhou, People’s Republic of China;(2) Key Laboratory for Biotechnology on Medicinal Plants of Jiangsu Province, Xuzhou Normal University, 221116 Xuzhou, People’s Republic of China;(3) College of Life Sciences, Nanjing Normal University, 210097 Nanjing, People’s Republic of China; |
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Abstract: | 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. |
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