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水稻HMGB1基因的克隆与特性分析

Cloning and characterization of rice HMGB1 gene.

作者信息

Wu Qiang, Zhang Wensheng, Pwee Keng-Hock, Kumar Prakash P

机构信息

Department of Biological Sciences, National University of Singapore, 10 Science Drive 4, Singapore 117543, Singapore.

出版信息

Gene. 2003 Jul 17;312:103-9. doi: 10.1016/s0378-1119(03)00605-x.

Abstract

We isolated a 918 bp long full-length rice HMGB1 cDNA, which has an open reading frame of 471 bp encoding 157 amino acids, with a central domain of high sequence similarity to the HMG-box domain of other plant HMGB1 proteins. RNA gel blot analysis indicated that rice HMGB1 gene is constitutively expressed in various tissues and organs. Southern hybridization and sequence analyses suggested that a single copy of the HMGB1 gene composed of seven exons and six introns exists in rice. We have also cloned a 1755 bp long 5' flanking region of the rice HMGB1 gene, which can be regarded as its promoter. 5' deletion analysis of this promoter indicated that positive cis-elements residing between -1400 and -1115 are important to enhance quantitative expression, whereas negative cis-elements between -1755 and -1400 and between -1115 and -351 inhibit expression.

摘要

我们分离出一个长度为918 bp的水稻HMGB1全长cDNA,其开放阅读框为471 bp,编码157个氨基酸,其中心结构域与其他植物HMGB1蛋白的HMG-box结构域具有高度序列相似性。RNA凝胶印迹分析表明,水稻HMGB1基因在各种组织和器官中组成型表达。Southern杂交和序列分析表明,水稻中存在由7个外显子和6个内含子组成的单拷贝HMGB1基因。我们还克隆了水稻HMGB1基因1755 bp长的5'侧翼区域,可将其视为启动子。对该启动子的5'缺失分析表明,位于-1400至-1115之间的正向顺式元件对于增强定量表达很重要,而位于-1755至-1400之间以及-1115至-351之间的负向顺式元件则抑制表达。

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