Goddijn O J, van der Duyn Schouten P M, Schilperoort R A, Hoge J H
Institute of Molecular Plant Sciences, Leiden University, Clusius Laboratory, Netherlands.
Plant Mol Biol. 1993 Aug;22(5):907-12. doi: 10.1007/BF00027376.
A novel selection system for plant genetic transformation was developed based on the enzyme tryptophan decarboxylase (TDC; EC 4.1.1.28) from Catharanthus roseus. This enzyme converts the toxic tryptophan analogue 4-methyl tryptophan (4-mT) into the non-toxic compound 4-methyl tryptamine. Expression of tdc in transgenic plants that have no endogenous TDC-activity allows selection on 4-mT. A vector was constructed containing a tdc cDNA clone under control of the constitutively expressed cauliflower mosaic virus 35S promoter. This vector was used in Agrobacterium-mediated tobacco leaf disc transformation experiments. The optimal concentration for selection with 4-mT was found to be 0.1 mM. The transformed nature of shoots obtained after tdc gene transfer and subsequent selection on 0.1 mM 4-mT was confirmed by northern blot analysis.
基于来自长春花的色氨酸脱羧酶（TDC；EC 4.1.1.28）开发了一种用于植物遗传转化的新型选择系统。该酶将有毒的色氨酸类似物4-甲基色氨酸（4-mT）转化为无毒化合物4-甲基色胺。在没有内源性TDC活性的转基因植物中tdc的表达允许在4-mT上进行选择。构建了一个载体，其中tdc cDNA克隆受组成型表达的花椰菜花叶病毒35S启动子控制。该载体用于农杆菌介导的烟草叶盘转化实验。发现用4-mT进行选择的最佳浓度为0.1 mM。通过Northern印迹分析证实了tdc基因转移并随后在0.1 mM 4-mT上进行选择后获得的芽的转化性质。
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