茅苍术DXS基因的克隆与分析

Cloning and Analysis of DXS Gene from Atractylodes lancea

目的:克隆茅苍术萜类化合物生物合成关键酶1-脱氧木糖-5-磷酸合酶(DXS)基因,并进行序列特征分析和组织特异性表达分析。方法:根据转录组测序所得的DXS基因片段,克隆出全长c DNA序列。运用实时荧光定量聚合酶链式反应(Real-time PCR),以tublin为内参基因,检测DXS基因在不同组织中的表达量。结果:克隆得到的DXS基因全长c DNA序列为2 151 bp,编码716个氨基酸,并在Gen Bank注册(登录号KY659208)。氨基酸序列系统发育分析表明,该序列与甜叶菊等菊科植物DXS基因有较高的同源性。Real-time PCR法检测发现茅苍术DXS在叶片中表达量最高,其次为花,而在根茎和根中表达很低。结论:获得了茅苍术DXS基因的全长c DNA序列,对其进行了初步生物信息学分析,揭示了其组织差异性表达特征,为进一步阐述该基因在茅苍术萜类成分生物合成途径中的功能奠定了基础。

Objective：This study is aimed to clone the key enzyme 1-deoxylated xylose-5-phosphate synthase（DXS） genes and analyze its tissue-specific expression in Atractylodes lancea.Method：The full-length c DNA sequence was cloned from the DXS gene fragment based on transcriptome sequencing data of A.lancea ge nerated in our previous study.The expression levels of DXS gene in different tissues of A.lancea were detected by Real-time PCR,with tublin as the reference gene.Result：The full-length c DNA sequence of DXS gene is 2 151 bp,encoding 716 amino acids,which has been deposited in Gen Bank（accession number KY659208）.Results of phylogenetic analysis showed that the sequence had high homology with DXS gene of other compositae plants,such as Stevia rebaudiana.Real-time PCR analysis showed that the DXS gene was mostly expressed in leaves,followed by flowers,rhizome and roots.Conclusion：Full-length c DNA sequence of DXS gene in A.lancea has been obtained for preliminary bioinformatics analysis,revealing differential expression in different tissues.The results will provide a groundwork for studying the function of DXS in terpenoid biosynthesis of A.lancea.