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拟南芥发育种子中FAE1基因的表达及FAE1启动子活性

Expression of the FAE1 gene and FAE1 promoter activity in developing seeds of Arabidopsis thaliana.

作者信息

Rossak M, Smith M, Kunst L

机构信息

Department of Botany, University of British Columbia, Vancouver, Canada.

出版信息

Plant Mol Biol. 2001 Aug;46(6):717-25. doi: 10.1023/a:1011603923889.

Abstract

Plant fatty acid elongase which catalyzes very-long-chain fatty acid (VLCFA) biosynthesis is a membrane-bound multienzyme complex. It is composed of four enzymes, a 3-ketoacyl-CoA synthase (condensing enzyme), a 3-ketoacyl-CoA reductase, a 3-hydroxyacyl-CoA dehydrase, and an enoyl-CoA reductase required for completion of each step of 2-carbon elongation of fatty acids. To improve our understanding of the overall regulation of the fatty acid elongase, we investigated the spatial and temporal expression of its key component, the FAE1-condensing enzyme, and examined the activity of the promoter of the FAE1 gene in Arabidopsis. In situ hybridization results revealed that FAE1 transcripts were found exclusively in the embryo. RNA blot analysis and histochemical analysis of GUS activity in pFAE1::GUS transgenic Arabidopsis lines demonstrated that the FAE1 gene was already transcribed in the early torpedo stage embryos 4-5 days after flowering, with transcription reaching its peak 9-11 days after flowering. VLCFA deposition closely paralleled FAE1 transcript accumulation. FAE1 promoter was highly active and embryo-specific. Because its timing coincides with the period of major storage lipid accumulation, and because its in vivo activity in Arabidopsis is superior to the napin promoter, FAE1 promoter may be ideal for genetic engineering of seed oil composition.

摘要

催化极长链脂肪酸(VLCFA)生物合成的植物脂肪酸延长酶是一种膜结合多酶复合体。它由四种酶组成,即3-酮酰基辅酶A合酶(缩合酶)、3-酮酰基辅酶A还原酶、3-羟基酰基辅酶A脱水酶和烯酰基辅酶A还原酶,这些酶是脂肪酸2碳延长各步骤完成所必需的。为了增进我们对脂肪酸延长酶整体调控的理解,我们研究了其关键组分FAE1-缩合酶的时空表达,并检测了拟南芥中FAE1基因启动子的活性。原位杂交结果显示,FAE1转录本仅在胚中发现。对pFAE1::GUS转基因拟南芥株系进行RNA印迹分析和GUS活性的组织化学分析表明,FAE1基因在开花后4 - 5天的鱼雷胚早期就已转录,在开花后9 - 11天转录达到峰值。VLCFA沉积与FAE1转录本积累密切平行。FAE1启动子具有高活性且具有胚特异性。由于其时间与主要储存脂质积累期一致,且其在拟南芥中的体内活性优于napin启动子,FAE1启动子可能是种子油成分基因工程的理想选择。

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