Cloning and expression analysis of kaurenoic acid oxidase gene in Tripterygium wilfordii

ZHANG Yi-feng1,2 SU Ping2 HU Tian-yuan1 ZHOU Jia-wei1 GUAN Hong-yu1,2 GAO Wei1,3 HUANG Lu-qi2

(1.School of Traditional Chinese Medicine, Capital Medical University, Beijing, China 100069)
(2.State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, Chinese Academy of Chinese Medical Sciences, Beijing, China 100700)
(3.Beijing Key Laboratory of Traditional Chinese Medicine Collateral Disease Theory Research, Beijing, China 100069)

【Abstract】Kaurenoic acid oxidase is involved in biosynthesis pathway of gibberellin. According to the transcriptome database, the specific primers were designed and used in cloning the full-length cDNA of TwKAO, the bioinformatic analysis of the sequence was performed. The qRT-PCR were used to detect the expression level of TwKAO after MeJA treatment. The full-length cDNA of the TwKAO was 1 874 bp encoding a polypeptide of 487 amino acids. The calculate molecular weight was about 56.02 k Da, and the theoretical isoelectric point(p I) was 8.89. The relative expression level of TwKAO was deduced by MeJA and reached the highest at 12 h after the treatment. Plant tissue expression analysis indicated that, TwKAO expressed the highest in leaves, while lowest in roots. For the first time, we cloned and analyzed the expression characteristics of TwKAO, which laid a foundation for deep analysis of growing development and terpenoid secondary metabolites in T. wilfordii.

【Keywords】 Tripterygium wilfordii; KAO; cloning; bioinformatics analysis; expression analysis;

【DOI】

【Funds】 Youth Project, National Natural Science Foundation of China (81422053); National Natural Science Foundation of China (81373906) Youth Elite Project, National Natural Science Foundation of China (81325023)

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(Translated by 梁兆晖)

    References

    [12]Ou C Q, Jiang S L, Wang F, et al. Cloning and Expression Analysis of ent-Kaurenoic Acid Oxidase Gene (PcKAO1)in Pear[J]. Acta Horticulturae Sinica, 2013, 40(5): 849-858 (in Chinese).

This Article

ISSN:1001-5302

CN: 11-2272/R

Vol 42, No. 01, Pages 88-93

January 2017

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Abstract

  • 1 Materials
  • 2 Method
  • 3 Results and analysis
  • 4 Discussion
  • References