茅苍术两个DXR基因(AlDXR)的克隆与分析

Cloning and Analysis of Two DXR Genes (AlDXR) in Atractylodes lancea

茅苍术萜类化合物生物合成关键酶1-脱氧-D-木酮糖5-磷酸还原酶异构化酶基因序列(AlDXR)尚未阐明,本研究克隆其基因序列并分析其序列特征,以期为阐明茅苍术萜类化合物生物合成分子机制提供帮助。从已获得的茅苍术转录组序列中筛选出DXR基因,克隆其全长cDNA序列,然后分析基因编码氨基酸序列特征、蛋白亚细胞定位、蛋白结构预测、系统发育等。成功筛选并克隆出两条全长cDNA序列分别为1 431 bp和1 422 bp的DXR基因(GenBank登录号分别为MG859908和MG859909),分别编码476个和473个氨基酸,系统发育分析表明二者分别与橡胶草和野菊花具有较高的同源性。本研究获得两个茅苍术DXR基因的全长cDNA序列,并对其进行初步生物信息学分析,为进一步阐明该基因在茅苍术萜类化合物生物合成中的功能提供指导。

The 1-Deoxy-D-xylulose 5-phosphate Reductase isomerase gene sequence(AlDXR), a key enzyme in terpenoids biosynthesis, has not been elucidated in Atractylodes lancea(Thunb.) DC. In this study, the AlDXR gene sequences were cloned and analyzed to offer help for molecular mechanism of terpenoid biosynthesis in A.lancea. We selected DXR gene from transcriptome sequence in obtained Atractylodes lancea, cloned full length cDNA sequence, and analyzed amino acid sequence characteristics in gene coding, protein subcellular localization,protein structure prediction and phylogeny. Two AlDXR genes were screened and cloned from transcriptome data and their 1 431 bp and 1 422 bp full-length cDNA sequences were obtained, which encode 476 and 473 amino acids, respectively(GenBank accession numbers MG859908 and MG859909). Phylogenetic analysis showed the highest homology with Taraxacum kok-saghyz and Chrysanthemum indicum var. aromaticum respectively. This study obtained two full-length cDNA sequence of DXR gene in Atractylodes lancea, and carried out preliminary bioinformatics analysis for it. These results will provide a guidance for the function of this gene in terpenoid biosynthesis in A. lancea.