天然截短型蛋白EsDREB2B的抗旱分子机理研究

项目名称： 天然截短型蛋白EsDREB2B的抗旱分子机理研究

项目编号： No.31500225

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 生物科学

项目作者： 李小双

作者单位： 中国科学院新疆生态与地理研究所

项目金额： 21万元

中文摘要： 天然截短型DREB蛋白是近年来新发现的一类特殊的、具有截短的C端结构的 DREB转录因子蛋白，目前对该类蛋白的研究较少，且其转录激活、转录调控和胁迫抗性提高的机理尚不明确且有待研究。EsDREB2B是申请者博士阶段从沙漠植物准噶尔无叶豆中克隆得到的一个新的天然截短型DREB蛋白，其虽是截断型蛋白，却有着广泛的胁迫抗性。本项目在明确了EsDREB2B抗逆功能的基础上，拟通过对其系列结构域缺失片段开展转录激活活性分析，明确它的转录激活域；通过对EsDREB2B系列结构域缺失片段在拟南芥中的抗旱功能比较，阐明其对抗旱性起关键作用的结构域；通过转录组分析野生型和过表达EsDREB2B基因拟南芥的差异表达基因，鉴定EsDREB2B所调控的下游靶基因；并通过酵母双杂交技术筛选与EsDREB2B互作的蛋白。通过上述研究揭示EsDREB2B的抗旱分子机理，并加深对天然截短型DREB抗逆应答机制的认识。

中文关键词： 转录因子；抗旱性；蛋白质相互作用；基因表达；干旱胁迫

英文摘要： Nature truncated DREB (Dehydration Response Element Binding protein) proteins are a kind of special proteins with truncated C-terminal, which are rarely reported and the exact mechanisms of the transcription activation, transcriptional regulation and stress tolerance are still unknown. As part of my dissertation research, a new naturally truncated DREB2-type protein (EsDREB2B) containing several important predicted domains was isolated from extreme drought-tolerant desert plant Eremosparton songoricum. EsDREB2B protein has showed strong and multiple stress tolerance, although truncated in gene structure. The aim of this proposed research is to reveal the molecular mechanism of stress tolerance of this truncated protein EsDREB2B. A series of deletion mutations of EsDREB2B gene will be constructed and analyzed. Specifically, 1) based on the transcriptional activation activity analysis in yeast system, the transcriptional activation domain of EsDREB2B are determined; 2) according to the drought tolerance performance of over-expression system in transgenic Arabidopsis, the stress tolerance-related domains of EsDREB2B are decided; 3) on the basis of the transcriptome comparison analysis between transgenic and wild type Arabidopsis, the target genes of EsDREB2B will be identified and 4) using yeast two-hybrid and BIFC analysis, the proteins interacted with EsDREB2B are confirmed. Through the above research, we aim to illustrate the molecular mechanism of the drought tolerances mediated by EsDREB2B gene. Our research may also helpful for understanding and revealing the new mechanism of truncated DREB proteins.

英文关键词： transcription factor;drought-resistance;protein interaction;gene expression;drought stress