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Soybean GH3 promoter contains multiple auxin-inducible elements.

机译:大豆GH3启动子包含多种生长素诱导型元素。

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摘要

The soybean GH3 gene is transcriptionally induced in a wide variety of tissues and organs within minutes after auxin application. To determine the sequence elements that confer auxin inducibility to the GH3 promoter, we used gel mobility shift assays, methylation interference, deletion analysis, linker scanning, site-directed mutagenesis, and gain-of-function analysis with a minimal cauliflower mosaic virus 35S promoter. We identified at least three sequence elements within the GH3 promoter that are auxin inducible and can function independently of one another. Two of these elements are found in a 76-bp fragment, and these consist of two independent 25- and 32-bp auxin-inducible elements. Both of these 25- and 32-bp auxin-inducible elements contain the sequence TGTCTC just upstream of an AATAAG. An additional auxin-inducible element was found upstream of the 76-bp auxin-inducible fragment; this can function independently of the 76-bp fragment. Two TGA-box or Hex-like elements (TGACGTAA and TGACGTGGC) in the promoter, which are strong binding sites for proteins in plant nuclear extracts, may also elevate the level of auxin inducibility of the GH3 promoter. The multiple auxin-inducible elements within the GH3 promoter contribute incrementally to the overall level of auxin induction observed with this promoter.
机译:施用生长素后数分钟内,大豆GH3基因在多种组织和器官中被转录诱导。为了确定赋予生长素诱导性的GH3启动子的序列元件,我们使用了最小的花椰菜花叶病毒35S启动子,进行了凝胶迁移率迁移测定,甲基化干扰,缺失分析,接头扫描,定点诱变和功能获得分析。 。我们确定了生长素诱导型GH3启动子内的至少三个序列元素,可以彼此独立地发挥作用。在一个76 bp的片段中发现了其中两个元素,它们由两个独立的25和32 bp生长素诱导型元素组成。这些25和32 bp的生长素诱导因子都在AATAAG的上游包含序列TGTCTC。在76 bp的生长素诱导片段的上游发现了另一种生长素诱导因子。它可以独立于76 bp片段起作用。启动子中的两个TGA-box或Hex-like元素(TGACGTAA和TGACGTGGC)是植物核提取物中蛋白质的强结合位点,也可以提高GH3启动子的生长素诱导性水平。 GH3启动子内的多种生长素诱导型元件对使用该启动子观察到的生长素诱导总体水平有增量贡献。

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