Expression of Strictosidine Synthase in Selected Subcellular Compartments of Tobacco Plant
异胡豆苷合成酶在烟草亚细胞区室的表达
查看参考文献16篇
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文摘
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Strictosidine synthase (STR) is a key enzyme involved in the biosynthesis of terpenoid indole al-kaloids (T1A) by condensing tryptamine and secologanin into strictosidine. The transgenic tobacco plants tar-geting STR to subcellular compartments were established to express STR in chloroplast, vacuole and endoplas-mic 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 enzy-matic assay was measured indirectly by lluorimetrically 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 ex-pressing 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 implica-tions on our understanding of the complex intracellular trafficking in metabolic intermediates of TIA biosynthesis. |
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其他语种文摘
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异胡豆苷合成酶（strictosidine synthase,STR）是吲哚生物碱生物合成的一种关键酶，将色胺（trypamine）和裂环马钱子（secologanin）耦合成为吲哚生物碱的前体化合物异胡豆苷。将异胡豆苷合成酶标定在烟草植物不同的亚细胞区室-叶绿林、液泡和内质网中表达，通过蛋白免疫印迹分析和STR酶活性的测定，表明STR在叶绿体，液泡和内质网中有效表达。STR体外酶活性分析采用间接荧光法检测色胺在反应体系的消耗。STR的酶活性分析表明了STR在烟草中不同的亚细胞区室得以活性表达。分离纯化转基因烟草的叶绿体，通过对其分离的不同部分的蛋白免疫印迹分析，确定了将STR正确标定在烟草的叶绿体中表达。 |
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来源
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Acta Botanica Sinica
,2002,44(5):579-582 【核心库】
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关键词
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Nicotiana tabacum
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strictosidine synthase
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subcellular compartment
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expression
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地址
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1.
Institute of Applied Ecology, The Chinese Academy of Sciences, Shenyang, 110016
2.
General Hospital in Shenyang Military District, Shenyang, 110016
3.
Botanical Institute of Rheinisch-Westfalisch Technische Hochschule Aachen, 德国, Aachen
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语种
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英文 |
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文献类型
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研究性论文 |
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ISSN
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1672-6650 |
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学科
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植物学 |
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基金
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中国科学院重大项目
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文献收藏号
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CSCD:1365238
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