
%0 Journal Article
%A Liao, Zhihua 
%A Tan, Qiumin 
%A Chai, Yourong 
%A Zuo, Kaijing 
%A Chen, Min 
%A Gong, Yifu 
%A Peng Wang, 
%A Pi, Yan 
%A Tan, Feng 
%A Sun, Xiaofen 
%A Tang, Kexuan 
%T Cloning and characterisation of the gene encoding HMG-CoA reductase from <emph type="2">Taxus media</emph> and its functional identification in yeast
%J Functional Plant Biology
%D 2004/01/22
%V 31
%N 1
%P 73-81
%K cloning, HMGR, mevalonate biosynthesis, Taxus media, yeast complementation.
%R https://doi.org/10.1071/FP03153
%U https://www.publish.csiro.au/paper/FP03153 
%X In plants, the first committed step in the pathway for biosynthesis of isoprenoids is catalysed by 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR, EC: 1.1.1.34). Here we report for the first time the cloning of a full-length cDNA encoding HMGR (Tm&#8211;HMGR) from a taxol-producing gymnosperm, Taxus media Rehder. The full-length cDNA of Tm&#8211;HMGR (GenBank accession number: AY277740) was 2307 base pairs (bp), with a 1791-bp open reading frame (ORF) encoding a 596-amino-acid polypeptide. Bioinformatic analysis revealed that Tm&#8211;HMGR contained two trans-membrane domains and a catalytic domain, and showed high homology to other plant HMGRs. Phylogenetic analysis indicated that Tm&#8211;HMGR was more ancient than other plant HMGRs. The structural modelling showed that Tm&#8211;HMGR had the typical spatial structure of HMGRs whose catalytic domains could be folded and divided into three spatial domains, L-domain, N-domain and S-domain. Southern blot analysis revealed that Tm&#8211;HMGR belonged to a small HMGR gene family. Northern blot analysis showed that Tm&#8211;HMGR was expressed in roots, stems and needles, with higher expression in stems and needles than in roots. Functional complementation of Tm&#8211;HMGR in a HMGR-deficient mutant yeast demonstrated that Tm&#8211;HMGR mediated the biosynthesis of mevalonate and provided the general precursor for taxol biosynthesis. 
