Cloning and Expression Analysis on 3-Hydroxy-3-methylglutaryl-coenzyme A Reductase from Aquilaria sinensis

Abstract

ObjectiveTo clone the full-length cDNA of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) from Aquilaria sinensis (AsHMGR1) and to analyze its expression profile in different tissues and in response to different treatments. HMGR is the first rate-limiting enzyme for sesquiterpene synthesis in the mevalonate pathway. MethodsRT-PCR and RACE were used to clone the full-length cDNA of HMGR from A. sinensis based on the conserved HMGR gene fragments. The bioinformatic analysis was performed on its nucleic acid and protein sequence. The expression profile of AsHMGR1 in different tissues and in response to different treatments was analyzed by quantitative RT-PCR. ResultsThe full-length AsHMGR1 cDNA was 2026 bp, containing a 1719 bp open reading frame which encoded a protein of 572 amino acids. Amino acid sequence homology alignment and phylogenetic analysis demonstrated that AsHMGR1 belonged to the HMGR gene family. The detection of tissue expression patterns showed that AsHMGR1 was mainly expressed in the stem, followed by roots and branches. AsHMGR1 could be stimulated by methyl jasmonate and H2O2 to varying degrees in a time-dependent manner. ConclusionThese data will provide a foundation for further investigation on AsHMGR1 functions and regulatory mechanisms in sesquiterpene synthesis in A. sinensis.