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Interaction of nuclear factors with upstream sequences of a lipid body membrane protein gene from carrot.

机译:核因子与胡萝卜脂质体膜蛋白基因上游序列的相互作用。

摘要

To study the regulation of gene expression during embryo development, we isolated a gene, DC 59, expressed in embryos but not in mature carrot plants. Sequence and S1 analysis showed that the gene was composed of one exon encoding a polypeptide of 19 kilodaltons and was highly homologous to the lipid body membrane protein gene L3 from maize. The plant hormone abscisic acid regulated the accumulation of DC 59 mRNA. To understand the mechanism of embryo-specific and hormonal regulation of DC 59, 5' DNA fragments were incubated with nuclear proteins. Two adjacent regions (from -706 to -235) interacted with nuclear extracts from embryos, resulting in the formation of four complexes (C1, C2, C3, and C4). Factors involved in the formation of the C3 and C4 complexes could be competed with sequences upstream of DC 8, a gene that is coordinately expressed with DC 59 during embryo development. DNase I footprinting analysis revealed that nuclear extracts from embryos bound to four AT-rich sequences, and the protected motifs within fragment V were located in the highly homologous upstream regions of DC 59 and DC 8 genes.
机译:为了研究胚胎发育过程中基因表达的调控,我们分离了一个在胚胎中表达但不在成熟胡萝卜植物中表达的基因DC 59。序列和S1分析表明,该基因由一个编码19道尔顿多肽的外显子组成,与玉米脂质体膜蛋白基因L3高度同源。植物激素脱落酸调节DC 59 mRNA的积累。为了了解DC 59的胚胎特异性和激素调节机制,将5'DNA片段与核蛋白一起孵育。两个相邻区域(从-706到-235)与胚胎的核提取物相互作用,导致形成四个复合物(C1,C2,C3和C4)。 C3和C4复合物的形成所涉及的因素可以与DC 8上游序列竞争,DC 8是在胚胎发育过程中与DC 59协同表达的基因。 DNase I足迹分析表明,与四个富AT序列结合的胚胎核提取物和片段V中受保护的基序位于DC 59和DC 8基因的高度同源上游区域。

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