丹参酮合成相关的候选基因CYP76AK5克隆及生物信息学分析

Cloning and Bioinformatic Analysisof CYP76AK5 Related to Tanshinone Biosynthesis in Salvia miltiorrhiza

目的:基于丹参基因组和转录组筛选、克隆及分析丹参酮合成途径候选的CYP450编码基因。方法:设计引物克隆SmCYP76AK5编码基因,通过MEGA软件构建系统进化树并结合实时定量PCR分析其在根不同组织(周皮、韧皮部、木质部)、根、茎、叶、花中的差异表达;最后利用在线生物信息学工具分析其理化性质,预测其二级结构、保守结构域等。结果:系统进化树分析表明SmCYP76AK5属于CYP71clan;基因差异表达显示SmCYP76AK5在根周皮中表达最高,与丹参酮的合成和积累相一致。结论:预测丹参SmCYP76AK5参与丹参酮的生物合成。

Objective：The candidate CYP450 gene related to tanshinone biosynthesis was selected and cloned based on genome-wide strategy in Salvia miltiorrhiza.Methods：The specific primers were designed to clone the SmCYP76AK5 gene.The phylogenetic tree was constructed by MEGA software.The different expression of SmCYP76AK5 in periderm，phloem，xylem，root，stem，leaf and flower was detected by real-time quantitative PCR.The physical and chemical properties，secondary structure，conserved domain were analyzed by a series of bioinformatic tools.Results：The phylogenetic analysis showed that SmCYP76AK5 belongs to CYP71clan.The expression of SmCYP76AK5 was the highest in periderm，in accordance with the synthesis and accumulation of tanshinone.Conclusion：SmCYP76AK5 was predicted to be involved in tanshinone production in S.miltiorrhiza.