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糖反应元件增强转基因烟草中马铃薯蛋白酶抑制剂II启动子的伤口反应。

Sugar response element enhances wound response of potato proteinase inhibitor II promoter in transgenic tobacco.

作者信息

Kim S R, Costa M A, An G H

机构信息

Institute of Biological Chemistry, Washington State University, Pullman 99164-6340.

出版信息

Plant Mol Biol. 1991 Nov;17(5):973-83. doi: 10.1007/BF00037137.

Abstract

The promoter region of the potato proteinase inhibitor II (PI-II) gene was studied to identify cis-acting regulatory sequences involved in sugar response using transgenic tobacco plants. The 5' control region covering an 892 nucleotide sequence upstream from the cap site and a 32 nucleotide untranslated region of the PI-II promoter was able to activate a reporter chloramphenicol acetyltransferase (cat) gene by wounding or by incubating in a sugar-free medium. This wound response was further enhanced by sugar. Hexoses, disaccharides, and some trisaccharides were strong inducers whereas pentoses, deoxy sugars, sugar acids, TCA cycle intermediates, amino acids, and other carbohydrates had little effect on the promoter activity. Deletion of the sequence between -892 and -573 abolished the wound response but not the sugar response. An additional 5' deletion to -453 removed the sugar inducibility. Locations of the cis-acting regulatory elements were further elucidated by 3' deletion analysis. Deletion of the downstream region from -520 did not affect the wound or sugar response of the promoter. However, 3' deletion mutant -574 was unable to respond to sugar but did respond weakly to wounding. Further deletion to -624 abolished both responses. Therefore, it can be concluded that a wound response element is located in between -624 and -574 and that the response is further enhanced by a sugar response element located in the sequence between -573 and -520.

摘要

利用转基因烟草植株研究了马铃薯蛋白酶抑制剂II(PI-II)基因的启动子区域,以鉴定参与糖响应的顺式作用调控序列。覆盖PI-II启动子帽位点上游892个核苷酸序列和32个核苷酸非翻译区的5'控制区,能够通过创伤或在无糖培养基中培养来激活报告氯霉素乙酰转移酶(cat)基因。糖进一步增强了这种创伤反应。己糖、二糖和一些三糖是强诱导剂,而戊糖、脱氧糖、糖酸、三羧酸循环中间产物、氨基酸和其他碳水化合物对启动子活性影响很小。-892至-573之间序列的缺失消除了创伤反应,但没有消除糖反应。进一步5'缺失至-453消除了糖诱导性。通过3'缺失分析进一步阐明了顺式作用调控元件的位置。从-520开始下游区域的缺失不影响启动子的创伤或糖反应。然而,3'缺失突变体-574对糖无反应,但对创伤反应较弱。进一步缺失至-624消除了两种反应。因此,可以得出结论,创伤反应元件位于-624和-574之间,并且该反应通过位于-573和-520之间序列中的糖反应元件进一步增强。

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