Morikami Atsushi, Matsunaga Rie, Tanaka Yoshimi, Suzuki Satomi, Mano Shoji, Nakamura Kenzo
Laboratory of Biochemistry, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya 464-8601, Japan.
Mol Genet Genomics. 2005 Feb;272(6):690-9. doi: 10.1007/s00438-004-1100-y. Epub 2005 Jan 15.
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.
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。