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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: To clone the gene 1-deoxy-xylulose-5-phosphate synthase (DXS), a key enzyme of terpenoid biosynthesis in Atractylodes lancea, and to analyze the sequence characteristics and tissue-specific expression. Methods: According to the DXS gene fragment obtained by transcriptome sequencing, the full-length c DNA sequence was cloned. Real-time PCR was used to detect the expression of DXS gene in different tissues using tublin as an internal control gene. RESULTS: The full-length c DNA sequence of the cloned DXS gene was 2 151 bp, encoding 716 amino acids and was registered at Gen Bank (accession number KY659208). Phylogenetic analysis of the amino acid sequence showed that this sequence shared high homology with the DXS gene of compositae such as Stevia. Real-time PCR assay showed that the highest expression level of DXS was found in leaves of A. officinalis, followed by flowers, but low in roots and roots. CONCLUSION: The full-length c DNA sequence of DXS gene of Atractylodes lancea has been obtained, and preliminary bioinformatics analysis has been carried out to reveal its differential expression characteristics. To further elucidate the biosynthesis pathway of terpenoids in Atractylodes lancea Function laid the foundation.