雷公藤贝壳杉烯酸氧化酶基因的全长cDNA克隆与表达分析

发布时间：2018-03-28 18:20

本文选题：雷公藤　切入点：贝壳杉烯酸氧化酶(KAO)　出处：《中国中药杂志》2017年01期

【摘要】：贝壳杉烯酸氧化酶(kaurenoic acid oxidase)是二萜赤霉素生物合成途径上的关键酶,参与植物生长发育等重要生物学过程。该文根据雷公藤转录组数据设计特异性引物,采用PCR技术克隆得到贝壳杉烯酸氧化酶全长c DNA序列,并进行生物信息学分析;使用实时定量PCR(qRT-PCR)研究基因的诱导表达水平。克隆得到Tw KAO长度为1 874 bp,编码487个氨基酸,蛋白相对分子质量56.02 k Da,理论等电点8.89;经茉莉酸甲酯(Me JA)诱导后,Tw KAO基因相对表达量在12 h达到峰值;经植株组织表达分析证实,Tw KAO基因在雷公藤的叶中表达量最高,根中最低。该研究首次克隆得到雷公藤KAO基因,并分析其mRNA表达特征,为深入研究雷公藤生长发育以及萜类活性成分次生代谢研究奠定基础。
[Abstract]:Kaurenoic acid oxidase is a key enzyme in the biosynthesis pathway of diterpenoid gibberellin, and is involved in plant growth and development. In this paper, specific primers were designed based on the transcriptional data of Tripterygium wilfordii. The full-length c DNA sequence of kapenoate oxidase was cloned by PCR technique and analyzed by bioinformatics. The induced expression level of the gene was studied by real-time quantitative PCR qRT-PCR.The length of Tw KAO was 1 874 BP, encoding 487 amino acids. The relative molecular weight of the protein was 56.02 kDa, the theoretical isoelectric point was 8.89, and the relative expression of Tw KAO gene reached its peak at 12 h after induction by methyl jasmonate (Me JA), and the highest expression of Tw KAO gene in the leaves of Tripterygium wilfordii was confirmed by plant tissue analysis. The KAO gene of Tripterygium wilfordii was cloned for the first time and its mRNA expression characteristics were analyzed, which laid a foundation for further study on the growth and development of Tripterygium wilfordii and the secondary metabolism of terpene active components.
【作者单位】：首都医科大学中医药学院;中国中医科学院中药资源中心道地药材国家重点实验室培育基地;中医病络研究北京市重点实验室;
【基金】：国家自然科学基金优秀青年基金项目(81422053);国家自然科学基金面上项目(81373906) 国家杰出青年科学基金项目(81325023)
【分类号】：S567.19

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