Characterization of the ethanol-inducible alc gene-expression system in Arabidopsis thaliana
Characterization of the ethanol-inducible alc gene-expression system in Arabidopsis thaliana
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DOI：
10.1046/j.1365-313x.2001.01146.x
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发表时间：
2001-10-01
期刊：
影响因子：
7.2
通讯作者：
Caddick, MX
中科院分区：
文献类型：
--
作者：
Roslan, HA;Salter, MG;Caddick, MX
植物中转基因的受控表达是基因功能表征和生长发育调控的关键。 alc基因表达系统源自丝状真菌构巢曲霉，此前已成功应用于烟草和马铃薯，并且具有在农业中应用的潜力。它对基础研究的价值很大程度上取决于它在拟南芥中的效用。我们对拟南芥的 ak 调节子进行了详细的功能分析。通过将 alcA 启动子与 β-葡萄糖醛酸酶 (GUS)、荧光素酶 (LUC) 和绿色荧光蛋白 (GFP) 基因连接，我们证明 alcR 介导的表达以高度响应的方式发生在整个植物中。诱导在一小时内发生，并且具有剂量依赖性，在没有外源诱导剂的情况下，土壤生长的植物的活性可以忽略不计。直接施用乙醇或将整个植物暴露在乙醇蒸气中是同样有效的诱导方法。使用土壤生长的植物的最大表达发生在诱导5天后。在大多数转基因中，表达受到严格调控且可逆。我们描述了在拟南芥中利用 ak 系统的最佳策略。
Controlled expression of transgenes in plants is key to the characterization of gene function and the regulated manipulation of growth and development. The alc gene-expression system, derived from the filamentous fungus Aspergillus nidulans, has previously been used successfully in both tobacco and potato, and has potential for use in agriculture. Its value to fundamental research is largely dependent on its utility in Arabidopsis thaliana. We have undertaken a detailed function analysis of the ak regulon in A. thaliana. By linking the alcA promoter to beta -glucuronidase (GUS), luciferase (LUC) and green fluorescent protein (GFP) genes, we demonstrate that alcR-mediated expression occurs throughout the plant in a highly responsive manner. Induction occurs within one hour and is dose-dependent, with negligible activity in the absence of the exogenous inducer for soil-grown plants. Direct application of ethanol or exposure of whole plants to ethanol vapour are equally effective means of induction. Maximal expression using soil-grown plants occurred after 5 days of induction. In the majority of transgenics, expression is tightly regulated and reversible. We describe optimal strategies for utilizing the ak system in A. thaliana.