Yun D J, Hashimoto T, Yamada Y
Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University, Japan.
Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):11799-803. doi: 10.1073/pnas.89.24.11799.
The tropane alkaloid scopolamine is a medicinally important anticholinergic drug present in several solanaceous plants. Hyoscyamine 6 beta-hydroxylase (EC 1.14.11.11) catalyzes the oxidative reactions in the biosynthetic pathway leading from hyoscyamine to scopolamine. We introduced the hydroxylase gene from Hyoscyamus niger under the control of the cauliflower mosaic virus 35S promoter into hyoscyamine-rich Atropa belladonna by the use of an Agrobacterium-mediated transformation system. A transgenic plant that constitutively and strongly expressed the transgene was selected, first by screening for kanamycin resistance and then by immunoscreening leaf samples with an antibody specific for the hydroxylase. In the primary transformant and its selfed progeny that inherited the transgene, the alkaloid contents of the leaf and stem were almost exclusively scopolamine. Such metabolically engineered plants should prove useful as breeding materials for obtaining improved medicinal components.
托烷生物碱东莨菪碱是一种重要的药用抗胆碱能药物，存在于多种茄科植物中。莨菪碱6β-羟化酶（EC 1.14.11.11）催化从莨菪碱到东莨菪碱的生物合成途径中的氧化反应。我们通过农杆菌介导的转化系统，将在花椰菜花叶病毒35S启动子控制下的来自黑莨菪的羟化酶基因导入富含莨菪碱的颠茄中。首先通过筛选卡那霉素抗性，然后用针对羟化酶的特异性抗体对叶片样品进行免疫筛选，选择出组成型且强表达转基因的转基因植物。在继承了转基因的初级转化体及其自交后代中，叶片和茎中的生物碱含量几乎全部是东莨菪碱。这种代谢工程植物应被证明可作为获得改良药用成分的育种材料。
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