Cloning, Bioinformatics Analysis and Expression Vector Construction of <I>DXS</I> Gene from <i>Aconitum vilmorinianum</i>

Abstract

<i>Aconitum vilmorinianum</i> is an important Chinese medicinal herb, which contains diterpenoid alkaloids and has the functions of antirheumatic activity, improvement in human meridians, relief of swelling and pain, activation of the blood circulation, and inhibition of tumor growth. The biosynthesis of diterpenoid alkaloids requires intermediate products obtained through terpenoid skeleton metabolism, in order to synthesize diterpenoid alkaloids. The synthesis of terpenoid skeletons includes the MVA pathway and the MEP pathway. 1-deoxyglucose-5-phosphate synthase (<I>DXS</I>) is the first rate limiting enzyme gene in the MEP pathway and plays an important regulatory role in the synthesis of terpenoids. Objective: To understand the characteristics of the DXS gene in Polygonummultiflorum. Method: By extracting RNA from the leaves of <i>A. vilmorinianum</i>and reverse transcribing it into cDNA, according to the transcriptome data of <i>A. vilmorinianum</i>, primers were designed to clone the full length of the gene, and the sequence of the coding protein was analyzed by bioinformatics. Construct overexpression vectors and inhibitory expression vectors. Results: The results showed that the coding region of the gene is 1512 bp, with 503 amino acids, molecular formula C₂₄₀₈H₃₈₂₂N₆₅₂O₇₁₃S₂₂, molecular weight 54020.10, the isoelectric point pI 6.29, the instability index 35.70, and belongs to the stable protein category. Successfully constructing overexpression vectors and inhibitory expression vectors can provide scientific basis for further research on the function of this gene in the diterpene biosynthesis pathway.