Cloning and analysis of a key enzyme of HMGR1 gene involved in terpene biosynthesis of Cornus officinalis / 中草药

ABSTRACT

Objective: To clone and analyze the cDNA sequence of 3-Hydroxy-3-methylglutaryl coenzyme-a reductase (HMGR1) of Cornus officinalis. Methods: In this study, specific primers were designed at both ends of the open reading frame (ORF) based on the unigene (c100572_g1) in the transcriptome data from C. officinalis. Subsequently, the cDNA sequence of CoHMGR1 gene was amplified by RT-PCR, cloned into the pTOPO-T vector and sequenced. This gene and its encoded protein were analyzed using bioinformatics methods. Results: The results suggested that CoHMGR1 was 2 116 bp in length andthe ORF was 1 338 bp in length, which encodes 445 amino acids and is a hydrophobic protein. Multiple sequence alignment and phylogenetic analysis showed that the amino acid sequence encoded by this gene has a high similarity with that of Camptotheca acuminate, reaching 79.56%. Based on the first comparison of transcriptome sequencing from leaves and fruits of C. officinalis, the CoHMGR1 gene was successfully cloned and analyzed. Conclusion: This study laid a foundation for studying the function of CoHMGR1 protein and the molecular mechanism of terpene biosynthesis pathway of C. officinalis.