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小麦组蛋白H3基因的近端启动子区域赋予转化水稻细胞S期特异性基因表达。

Proximal promoter region of the wheat histone H3 gene confers S phase-specific gene expression in transformed rice cells.

作者信息

Ohtsubo N, Nakayama T, Terada R, Shimamoto K, Iwabuchi M

机构信息

Department of Botany, Faculty of Science, Kyoto University, Japan.

出版信息

Plant Mol Biol. 1993 Nov;23(3):553-65. doi: 10.1007/BF00019303.

Abstract

The cis-regulatory elements that confer cell cycle-dependent expression to the wheat histone H3 gene were investigated in rice cells (Oc strain) transformed with H3/GUS chimeric genes. 5' deletion mutants of the H3 promoter region (from -1711, -908 or -185 to +57 relative to the transcription start site) were joined to the coding sequence of the bacterial beta-glucuronidase (GUS) gene then introduced stably into rice cells. S1 analyses of the RNA from transformed rice cells whose cell cycles had been synchronized by treatment with aphidicolin showed that the steady-state levels of the transcripts from chimeric genes were altered with the change in DNA synthesis and the content of rice H3 mRNA throughout the cell cycle. Even though H3 promoter activity decreased as 5' deletion proceeded, transcripts from the chimeric genes showed increases, as much as 10-fold 1 h after release from the aphidicolin block, which were rapidly lost over the next 4 h. The results suggest that the 242 bp sequence from -185 to +57, which contains the basal promoter region, confers the S phase-specific expression of the H3 gene and that the upstream sequence from position -186 is required for the full activity of this promoter.

摘要

利用H3/GUS嵌合基因转化的水稻细胞(Oc株)研究了赋予小麦组蛋白H3基因细胞周期依赖性表达的顺式调控元件。将H3启动子区域的5'缺失突变体(相对于转录起始位点从-1711、-908或-185至+57)与细菌β-葡萄糖醛酸酶(GUS)基因的编码序列连接,然后稳定导入水稻细胞。对经抑蚜灵处理使细胞周期同步化的转化水稻细胞的RNA进行S1分析表明,嵌合基因转录本的稳态水平随DNA合成的变化以及整个细胞周期中水稻H3 mRNA含量的变化而改变。尽管随着5'缺失的进行H3启动子活性降低,但嵌合基因的转录本在从抑蚜灵阻断中释放1小时后显示出增加,高达10倍,随后在接下来的4小时内迅速消失。结果表明,包含基础启动子区域的从-185至+57的242 bp序列赋予了H3基因的S期特异性表达,并且-186位上游序列是该启动子充分活性所必需的。

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