Expression of strictosidine synthase in selected subcellular compartments of tobacco plant

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	Expression of strictosidine synthase in selected subcellular compartments of tobacco plant
其他题名	Expression of strictosidine synthase in selected subcellular compartments of tobacco plant
	Wang M 1; Li QR 1; Di Fiore S 1; Fischer R 1
	2002
发表期刊	ACTA BOTANICA SINICA
ISSN	0577-7496
卷号	44 期号:5 页码:579-582
摘要	Strictosidine synthase (STR) is a key enzyme involved in the biosynthesis of terpenoid indole alkaloids (TIA) by condensing tryptamine and secologanin into strictosidine. The transgenic tobacco plants targeting STR to subcellular Compartments were established to express STR in chloroplast, vacuole and endoplasmic reticulum (ER) by the,tobacco stable transformation. It was shown that STR was effectively expressed in the above subcellular Compartments by Western blot analysis and STR enzymatic assay. In vitro, STR enzymatic assay was measured indirectly by fluorimetrically detecting depletion of tryptamine feeding on secologanin in the reaction mixture. The tryptamine were completely depleted by STR in the crude extract of leaves of transgenic tobacco plants targeting and expressing STR in the chloroplast, vacuole and ER, which ascertained the STR functionally targeted to the three subcellular Compartments. To confirm STR correct targeting and expressing in chloroplast, the. chloroplasts were isolated and the fractions of purified chloroplasts were analyzed by Western blot. The hypothesis of STR correct targeting to the chloroplast was tested. The results have implications on our understanding of the complex intracellular trafficking in metabolic intermediates of TIA biosynthesis.
其他摘要	Strictosidine synthase (STR) is a key enzyme involved in the biosynthesis of terpenoid indole alkaloids (TIA) by condensing tryptamine and secologanin into strictosidine. The transgenic tobacco plants targeting STR to subcellular Compartments were established to express STR in chloroplast, vacuole and endoplasmic reticulum (ER) by the,tobacco stable transformation. It was shown that STR was effectively expressed in the above subcellular Compartments by Western blot analysis and STR enzymatic assay. In vitro, STR enzymatic assay was measured indirectly by fluorimetrically detecting depletion of tryptamine feeding on secologanin in the reaction mixture. The tryptamine were completely depleted by STR in the crude extract of leaves of transgenic tobacco plants targeting and expressing STR in the chloroplast, vacuole and ER, which ascertained the STR functionally targeted to the three subcellular Compartments. To confirm STR correct targeting and expressing in chloroplast, the. chloroplasts were isolated and the fractions of purified chloroplasts were analyzed by Western blot. The hypothesis of STR correct targeting to the chloroplast was tested. The results have implications on our understanding of the complex intracellular trafficking in metabolic intermediates of TIA biosynthesis.
关键词	TRYPTOPHAN DECARBOXYLASE Nicotiana tabacum strictosidine synthase subcellular compartment expression
收录类别	CSCD
语种	英语
CSCD记录号	CSCD:1365238
引用统计	被引频次：1[CSCD] [CSCD记录]
文献类型	期刊论文
条目标识符	http://ir.imr.ac.cn/handle/321006/148675
专题	中国科学院金属研究所
作者单位	1.中国科学院 2.中国科学院金属研究所 3.Rhein Westfal TH Aachen
推荐引用方式 GB/T 7714	Wang M,Li QR,Di Fiore S,et al. Expression of strictosidine synthase in selected subcellular compartments of tobacco plant[J]. ACTA BOTANICA SINICA,2002,44(5):579-582.
APA	Wang M,Li QR,Di Fiore S,&Fischer R.(2002).Expression of strictosidine synthase in selected subcellular compartments of tobacco plant.ACTA BOTANICA SINICA,44(5),579-582.
MLA	Wang M,et al."Expression of strictosidine synthase in selected subcellular compartments of tobacco plant".ACTA BOTANICA SINICA 44.5(2002):579-582.