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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。
Kaurenoic acid oxidase is a key enzyme in the biosynthetic pathway of terbanopyranil and participates in important biological processes such as plant growth and development. In this paper, specific primers were designed according to the data of Tripterygium translater. The full-length c DNA sequence of cedryl succinate oxidase was cloned by PCR and analyzed by bioinformatics. The induction of gene by qRT-PCR The expression level. The length of Tw KAO gene was 1 874 bp, encoding a protein of 487 amino acids. The molecular weight of the protein was 56.02 kDa and the theoretical isoelectric point was 8.89. The relative expression level of Tw KAO gene after induced by methyl jasmonate (Me JA) Reached the peak. The expression of Tw KAO gene was the highest in the leaves of Tripterygium wilfordii, and the lowest in the roots. In this study, we first cloned the KAO gene of Tripterygium wilfordii and analyzed its mRNA expression characteristics, which lays the foundation for the further study on the growth and development of Tripterygium wilfordii and the secondary metabolites of terpenoids.