3-Hydroxy-3-methylglutaryl-coenzymeA reductase (HMGR), the first rate-limiting enzyme of Mevalonate (MVA) pathway was isolated from Andrographis paniculata (ApHMGR) and expressed in bacterial cells. Full length ApHMGR (1937 bp) was submitted to NCBI with accession number MG271748.1. The open reading frame (ORF) was flanked by a 31-bp 5'-UTR, 118-bp 3'-UTR and ApHMGR contained a 1787 bp ORF encoding protein of 595 amino acids. ApHMGR protein was approximately 64 kDa, with isoelectric point of 5.75. Isolated ApHMGR was cloned into pET102 vector and expressed in E. coli BL21 (DE 3) cells, and characterized by SDS-PAGE. HPLC analysis for andrographolide content in leaf, stem and root of A. paniculata revealed highest in leaf tissue. The expression patterns of ApHMGR in different plant tissues using qRT-PCR revealed high in root tissue correlating with HPLC data. Three dimensional (3D) structural model of ApHMGR displayed 90% of the amino acids in most favored regions of the Ramachandran plot with 93% overall quality factor. ApHMGR was highly conserved with plant specific N-terminal membrane domains and C-terminal catalytic regions. Phylogenetic analysis showed A. paniculata sharing common ancestor with Handroanthus impetiginosus. 3D model of ApHMGR was screened for the interaction with substrates NADPH, HMG CoA and inhibitor using Auto Dock Vina. In silico analysis revealed that full length ApHMGR had extensive similarities to other plant HMGRs. The present communication reports the isolation of full length HMGR from A. paniculata, its heterologous expression in bacterial cells and in silico structural and functional characterization providing valuable genomic information for future molecular interventions.

中文翻译：

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穿心莲（Burm.f）Nees的3-羟基-3-甲基戊二酰辅酶A还原酶（HMGR）基因的分离，表征和计算机分析。

从穿心莲（ApHMGR）中分离出甲羟戊酸（MVA）途径的第一个限速酶3-羟基3-甲基戊二酰辅酶A（HMGR），并在细菌细胞中表达。全长ApHMGR（1937 bp）已提交NCBI，登录号MG271748.1。开放阅读框（ORF）的侧翼是31-bp的5'-UTR，118-bp的3'-UTR，并且ApHMGR含有1595bp的ORF编码595个氨基酸的蛋白质。ApHMGR蛋白约为64 kDa，等电点为5.75。将分离的ApHMGR克隆到pET102载体中，并在大肠杆菌BL21（DE 3）细胞中表达，并通过SDS-PAGE表征。高效液相色谱分析表明，穿心莲中叶，茎和根中的穿心莲内酯含量最高。使用qRT-PCR在不同植物组织中ApHMGR的表达模式表明，根部组织中的含量与HPLC数据相关。ApHMGR的三维（3D）结构模型显示了Ramachandran图最喜欢的区域中90％的氨基酸，总质量因子为93％。ApHMGR与植物特有的N端膜结构域和C端催化区域高度保守。系统发育分析表明，A。paniculata与Handroanthus impetiginosus共有共同祖先。使用Auto Dock Vina筛选了ApHMGR的3D模型与底物NADPH，HMG CoA和抑制剂的相互作用。电脑分析表明，全长ApHMGR与其他植物HMGR具有广泛的相似性。本来文报告了全长HMGR与拟南芥的分离，