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两个顺式作用调控元件参与了转基因烟草中甘薯sporamin基因启动子的蔗糖诱导表达。

Two cis-acting regulatory elements are involved in the sucrose-inducible expression of the sporamin gene promoter from sweet potato in transgenic tobacco.

作者信息

Morikami Atsushi, Matsunaga Rie, Tanaka Yoshimi, Suzuki Satomi, Mano Shoji, Nakamura Kenzo

机构信息

Laboratory of Biochemistry, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya 464-8601, Japan.

出版信息

Mol Genet Genomics. 2005 Feb;272(6):690-9. doi: 10.1007/s00438-004-1100-y. Epub 2005 Jan 15.

Abstract

In this study, we generated transgenic tobacco plants that express the beta-glucuronidase (GUS) gene under the control of the 305-bp 5'-upstream region of a gene coding for sporamin A of sweet potato. Expression of GUS in excised tobacco leaves was induced by sucrose, mimicking the sugar-inducible expression of the endogenous sporamin genes in sweet potato. Deletion of the sequences extending from position -305 (relative to the transcription start site) to -283 and from -146 to -87 resulted in an approximately 40-fold enhancement in GUS reporter expression. Furthermore, the sequence from -282 to -165 conferred sucrose-inducibility on the -89 core promoter of the Cauliflower Mosaic Virus 35S RNA gene. Analysis of internal deletions, linker scanning and the introduction of base substitutions in the sequence between positions -282 and -165 indicated that sucrose-responsiveness conferred by this region was dependent on the presence of two cis-acting elements, termed CMSREs (carbohydrate metabolite signal responsive elements) 1 and 2, which are separated by a spacer. A sequence similar or identical to the core of CMSRE-1 (TGGACGG) is also present in the promoters of several other sugar-inducible genes, and sequences encopassing the TGGACGG-related motifs from two of these could functionally replace the CMSRE-1 in the truncated sporamin A promoter. These results suggest that the TGGACGG element plays an important role in the sucrose-inducible expression of a group of plant genes.

摘要

在本研究中,我们培育了转基因烟草植株,这些植株在甘薯sporamin A编码基因305 bp的5'上游区域的控制下表达β-葡萄糖醛酸酶(GUS)基因。蔗糖可诱导切除的烟草叶片中GUS的表达,这模拟了甘薯中内源性sporamin基因的糖诱导表达。从位置-305(相对于转录起始位点)到-283以及从-146到-87的序列缺失导致GUS报告基因表达提高了约40倍。此外,从-282到-165的序列赋予了花椰菜花叶病毒35S RNA基因-89核心启动子蔗糖诱导性。对-282至-165之间序列的内部缺失、接头扫描和碱基替换分析表明,该区域赋予的蔗糖反应性取决于两个顺式作用元件的存在,这两个元件称为碳水化合物代谢物信号反应元件(CMSREs)1和2,它们被一个间隔区隔开。CMSRE-1(TGGACGG)核心的相似或相同序列也存在于其他几个糖诱导基因的启动子中,并且来自其中两个基因的包含TGGACGG相关基序的序列可以在截短的sporamin A启动子中功能性地替代CMSRE-1。这些结果表明,TGGACGG元件在一组植物基因的蔗糖诱导表达中起重要作用。

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