来自红树植物秋茄（Kandelia candel (L.) Druce）的多功能三萜合酶cDNA的分子克隆与功能表达
Molecular cloning and functional expression of a multifunctional triterpene synthase cDNA from a mangrove species Kandelia candel (L.) Druce.
作者信息
Basyuni Mohammad, Oku Hirosuke, Inafuku Masashi, Baba Shigeyuki, Iwasaki Hironori, Oshiro Keichiro, Okabe Takafumi, Shibuya Masaaki, Ebizuka Yutaka
机构信息
United Graduate School of Agricultural Sciences, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.
出版信息
Phytochemistry. 2006 Dec;67(23):2517-24. doi: 10.1016/j.phytochem.2006.09.016. Epub 2006 Oct 31.
Homology based PCRs with degenerate primers designed from the conserved sequences among the known oxidosqualene cylases (OSCs) have resulted in cloning of a triterpene synthase (KcMS) from the young roots of Kandelia candel (L.) Druce (Rhizophoraceae). KcMS consists of a 2286 bp open reading frame, which codes for 761 amino acids. The deduced amino acid sequence showed 79% homology to a lupeol synthase from Ricinus communis suggesting it to be a lupeol synthase of K. candel. KcMS was expressed in a lanosterol synthase deficient yeast with the expression vector pYES2 under the control of GAL1 promoter. GC-MS analysis showed that the transformant accumulated a mixture of lupeol, beta-amyrin and alpha-amyrin in a 2:1:1 ratio, indicating that KcMS encodes a multifunctional triterpene synthase, although it showed high sequence homology to a R. communis lupeol synthase. This is the first OSC cloning from mangrove tree species.
利用根据已知氧化鲨烯环化酶（OSC）的保守序列设计的简并引物进行基于同源性的聚合酶链式反应（PCR），已从秋茄（Kandelia candel (L.) Druce，红树科）的幼根中克隆出一种三萜合酶（KcMS）。KcMS由一个2286 bp的开放阅读框组成，编码761个氨基酸。推导的氨基酸序列与蓖麻（Ricinus communis）的羽扇豆醇合酶有79%的同源性，表明它是秋茄的羽扇豆醇合酶。KcMS在带有表达载体pYES2的羊毛甾醇合酶缺陷型酵母中，在GAL1启动子的控制下表达。气相色谱-质谱联用（GC-MS）分析表明，转化体积累了羽扇豆醇、β-香树脂醇和α-香树脂醇的混合物，比例为2:1:1，这表明KcMS编码一种多功能三萜合酶，尽管它与蓖麻羽扇豆醇合酶具有高度的序列同源性。这是首次从红树树种中克隆出OSC。
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