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Expression of strictosidine synthase in selected subcellular compartments of tobacco plant
Alternative TitleExpression of strictosidine synthase in selected subcellular compartments of tobacco plant
Wang M1; Li QR1; Di Fiore S1; Fischer R1
2002
Source PublicationACTA BOTANICA SINICA
ISSN0577-7496
Volume44Issue:5Pages:579-582
AbstractStrictosidine 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.
Other AbstractStrictosidine 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.
KeywordTRYPTOPHAN DECARBOXYLASE Nicotiana tabacum strictosidine synthase subcellular compartment expression
Indexed ByCSCD
Language英语
CSCD IDCSCD:1365238
Citation statistics
Cited Times:1[CSCD]   [CSCD Record]
Document Type期刊论文
Identifierhttp://ir.imr.ac.cn/handle/321006/148675
Collection中国科学院金属研究所
Affiliation1.中国科学院
2.中国科学院金属研究所
3.Rhein Westfal TH Aachen
Recommended Citation
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.
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