Hallard D, van der Heijden R, Verpoorte R, Cardoso M I Lopes, Pasquali G, Memelink J, Hoge J H C
Division of Pharmacognosy, Leiden/ Amsterdam Center for Drug Research, Leiden University, Gorlaeus Laboratories, PO Box 9502, 2300 RA Leiden, The Netherlands Fax no.: +31-71-527-45-11 e-mail:
Institute of Molecular Plant Sciences, Leiden University, Clusius Laboratory, PO Box 9505, 2300 RA Leiden, The Netherlands, , , , , , NL.
Plant Cell Rep. 1997 Nov;17(1):50-54. doi: 10.1007/s002990050350.
A transgenic cell suspension culture of Nicotiana tabacum L. `Petit Havana' SR1 was established expressing tryptophan decarboxylase and strictosidine synthase cDNA clones from Catharanthus roseus (L.) G. Don under the direction of cauliflower mosaic virus 35S promoter and nopaline synthase terminator sequences. During a growth cycle, the transgenic tobacco cells showed relatively constant tryptophan decarboxylase activity and an about two- to sixfold higher strictosidine synthase activity, enzyme activities not detectable in untransformed tobacco cells. The transgenic culture accumulated tryptamine and produced strictosidine upon feeding of secologanin, demonstrating the in vivo functionality of the two transgene-encoded enzymes. The accumulation of strictosidine, which occurred predominantly in the medium, could be enhanced by feeding both secologanin and tryptamine. No strictosidine synthase activity was detected in the medium, indicating the involvement of secologanin uptake and strictosidine release by the cells.
在花椰菜花叶病毒35S启动子和胭脂碱合酶终止子序列的调控下，建立了表达来自长春花（Catharanthus roseus (L.) G. Don）色氨酸脱羧酶和异胡豆苷合酶cDNA克隆的烟草（Nicotiana tabacum L. `Petit Havana' SR1）转基因细胞悬浮培养物。在一个生长周期中，转基因烟草细胞显示出色氨酸脱羧酶活性相对恒定，而异胡豆苷合酶活性高出约2至6倍，在未转化的烟草细胞中未检测到这些酶的活性。转基因培养物积累了色胺，并在添加番木鳖苷后产生了异胡豆苷，证明了这两种转基因编码酶在体内的功能。主要在培养基中发生的异胡豆苷积累可以通过同时添加番木鳖苷和色胺来增强。在培养基中未检测到异胡豆苷合酶活性，表明细胞参与了番木鳖苷的摄取和异胡豆苷的释放。
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