Molecular cloning, characterization and function analysis of the gene encoding HMG-CoA reductase from Euphorbia Pekinensis Rupr.
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.
School of Chemical Engineering, China University of Mining and Technology, Xuzhou, People's Republic of China.
Xiaoying
X
Cao
Zhimin
Z
Zong
Xiuyun
X
Ju
Yong
Y
Sun
Chuanchao
C
Dai
Qun
Q
Liu
Jihong
J
Jiang
D001483:Base Sequence; D015139:Blotting, Southern; D002338:Carotenoids / Q000096:biosynthesis; D003001:Cloning, Molecular; D019295:Computational Biology; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D028481:Euphorbia / Q000201:enzymology*; D020869:Gene Expression Profiling; D006903:Hydroxymethylglutaryl CoA Reductases / Q000235:genetics* / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000737:chemistry; D018517:Plant Roots / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
D017931:DNA Primers; D018076:DNA, Complementary; D002338:Carotenoids; D006903:Hydroxymethylglutaryl CoA Reductases