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通过对芥菜(Brassica juncea)谷胱甘肽S-转移酶基因启动子的5'-缺失分析,对其在拟南芥中的分子特征进行研究。

Molecular characterization of a Phi-class mustard (Brassica juncea) glutathione S-transferase gene in Arabidopsis thaliana by 5'-deletion analysis of its promoter.

作者信息

Gong Haibiao, Hu Wen-Wei, Jiao Yuxia, Pua Eng-Chong

机构信息

Plant Genetic Engineering Laboratory, Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Republic of Singapore.

出版信息

Plant Cell Rep. 2005 Sep;24(7):439-47. doi: 10.1007/s00299-005-0961-9. Epub 2005 May 31.

Abstract

Glutathione S-transferases (GSTs) are regulated by various stimuli at the transcriptional level. In this study, a 2,640-bp promoter sequence of a mustard GST gene, BjGSTF2, was cloned. Several truncated BjGSTF2 promoters were generated by 5'-deletion, fused to the beta-glucuronidase (GUS) coding sequence and the chimeric genes expressed in Arabidopsis thaliana. Transgene expression in GST2623::GUS plants carrying the longest promoter varied considerably. GUS activity was high in the roots, cotyledons, anthers and both ends of the silique, but it was low or barely detectable in the leaves, seeds, petals and stamens. Analysis of transgenic plants expressing the GUS gene under the control of different truncated BjGSTF2 promoters revealed several regions that possessed cis-acting elements required for the basal and induced expression by H(2)O(2), salicylic acid and 1-aminocyclopropane-1-carboxylate and down-regulation by spermidine. The results also showed that the GUS activity of GST2623::GUS coincided well with the H(2)O(2) accumulation pattern in cultured leaf-disc explants during the regeneration process.

摘要

谷胱甘肽S-转移酶(GSTs)在转录水平上受到多种刺激的调控。在本研究中,克隆了芥菜GST基因BjGSTF2的一段2640 bp的启动子序列。通过5'端缺失产生了几个截短的BjGSTF2启动子,将其与β-葡萄糖醛酸酶(GUS)编码序列融合,并在拟南芥中表达嵌合基因。携带最长启动子的GST2623::GUS植株中的转基因表达差异很大。GUS活性在根、子叶、花药和角果两端较高,但在叶、种子、花瓣和雄蕊中较低或几乎检测不到。对在不同截短的BjGSTF2启动子控制下表达GUS基因的转基因植株的分析揭示了几个区域,这些区域具有基础表达以及受H(2)O(2)、水杨酸和1-氨基环丙烷-1-羧酸诱导表达和受亚精胺下调所需的顺式作用元件。结果还表明,在再生过程中,GST2623::GUS的GUS活性与培养的叶盘外植体中H(2)O(2)积累模式吻合良好。

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