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穿心莲甲羟戊酸二磷酸脱羧酶基因的克隆与组织表达

[Cloning and tissue expression of mevalonate disphosphate decarboxylase genes from Andrographis paniculata].

作者信息

Wang Xiao-Yun, Chen Rong, Zhang En-Hui, Zhong Guo-Yue

机构信息

Collaborative Innovation Center for the Modern Technology and Industrial Development of Jiangxi Minority Traditional Medicine, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China.

Chinese Medicine Germplasm Resource Engineering Technology Research Center of Jiangxi Province, Nanchang 330004, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2016 Feb;41(4):636-642. doi: 10.4268/cjcmm20160415.

Abstract

Andrographolide is a main bioactive substance in Andrographis paniculata, and extensively used in anti-inflammatory drugs. In order to increase andrographolide production in plant, three 1260 bp ORFs encoding mevalonate disphosphate decarboxylases with 419 amino acids were cloned from A. paniculata by RACE method and analyzed by bioinformatic software. Their tissue expression patterns were predicted by real time PCR. Eleven conserved amino acid residues determining specificity and activity of these MVDs were predicted in these amino acid sequences, but no plastid targeted signal peptides were detected. These MVDs have high similarities with the MVD protein (GenBank number: AEZ55675.1) from Salvia miltiorrhiza. In stems and leaves, expression levels of these MVD genes were constant, and reached the highest level at bud stage and the beginning of flowering. The MVD genes we have cloned from A. paniculata could be used in genetic engineering of andrographolide biosynthsis pathway in future.

摘要

穿心莲内酯是穿心莲中的一种主要生物活性物质,广泛应用于抗炎药物中。为了提高植物中穿心莲内酯的产量,通过RACE方法从穿心莲中克隆了3个编码甲羟戊酸二磷酸脱羧酶的1260 bp开放阅读框,其含有419个氨基酸,并使用生物信息学软件进行分析。通过实时PCR预测它们的组织表达模式。在这些氨基酸序列中预测到11个决定这些甲羟戊酸二磷酸脱羧酶特异性和活性的保守氨基酸残基,但未检测到质体靶向信号肽。这些甲羟戊酸二磷酸脱羧酶与来自丹参的甲羟戊酸二磷酸脱羧酶蛋白(GenBank编号:AEZ55675.1)具有高度相似性。在茎和叶中,这些甲羟戊酸二磷酸脱羧酶基因的表达水平恒定,在芽期和开花初期达到最高水平。我们从穿心莲中克隆的甲羟戊酸二磷酸脱羧酶基因未来可用于穿心莲内酯生物合成途径的基因工程。

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