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拟南芥QRAP2基因的克隆、表达、DNA结合能力及体外转录活性分析 被引量:3
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作者 魏刚 雷娟 +1 位作者 巩威 朱玉贤 《科学通报》 EI CAS CSCD 北大核心 2005年第17期1863-1868,共6页
通过对拟南芥ATH1基因芯片数据的分析,从拟南芥中鉴定到一个受干旱胁迫高水平诱导的cDNA片段,并用RACE方法获得了其全长cDNA.PCR及定量实时PCR分析表明,该基因在经干旱、紫外线照射、脱落酸、高盐以及水杨酸等胁迫条件处理后表达量均有... 通过对拟南芥ATH1基因芯片数据的分析,从拟南芥中鉴定到一个受干旱胁迫高水平诱导的cDNA片段,并用RACE方法获得了其全长cDNA.PCR及定量实时PCR分析表明,该基因在经干旱、紫外线照射、脱落酸、高盐以及水杨酸等胁迫条件处理后表达量均有显著提高,特别是干旱处理后短时间内表达量迅速提高,3h后即提高到对照样品的430多倍.通过多序列比对和系统进化分析,我们将该基因归于DREB亚家族.由于该基因编码蛋白含有典型的AP2/EREBPDNA结合结构域,并在N端有一段富含谷氨酰氨残基的区域,所以将它命名为QRAP2(Glutamine-richAP2).凝胶阻滞实验结果显示,QRAP2蛋白能够特异结合DRE顺式元件序列但不能与mDRE和GCC等非相关元件结合.酵母单杂交实验表明,QRAP2蛋白全长序列或其N端112个氨基酸与GAL4DNA结合结构域形成的融合蛋白表现出转录激活功能,而其C端135个氨基酸与GAL4DNA结合结构域的融合蛋白则没有表现出转录激活活性.现有资料的分析表明,QRAP2是AP2/EREBP转录因子家族的新成员,可能在干旱胁迫条件下参与激活相关下游基因的表达. 展开更多
关键词 qrap2 胁迫 DREB 酵母单杂交 转录因子 CDNA片段 基因芯片 体外转录 拟南芥 活性分析
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Molecular cloning,expressional profiling,DNA binding and trans-activation property studies of QRAP2 from Arabidopsis thaliana 被引量:3
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作者 WEI Gang LEI Juan +1 位作者 GONG Wei ZHU Yuxian 《Chinese Science Bulletin》 SCIE EI CAS 2005年第17期1873-1878,共6页
Based upon analysis of data obtained from theATH1 microarrays, a cDNA that was highly induced afterdrought treatment, was isolated from Arabidopsis seedlings.RT-PCR and Quantitative Real-Time (QRT)-PCR experi-ments sh... Based upon analysis of data obtained from theATH1 microarrays, a cDNA that was highly induced afterdrought treatment, was isolated from Arabidopsis seedlings.RT-PCR and Quantitative Real-Time (QRT)-PCR experi-ments showed that expression level of the gene increasedsignificantly upon drought, UV, abscisic acid, high salinityand salicylic acid treatments. It was classified as a DREBsubfamily member based on multiple sequence alignmentand phylogenetic characterization. Since it encoded a proteinwith a typical ERF/AP2 DNA-binding domain and containeda glutamine-rich region near its N terminus, we named itQRAP2 (for glutamine-rich AP2). Gel retardation assay re-vealed that QRAP2 was able to form a specific complex withthe previously characterized DRE element while did notshow any affinity to the GCC box or the mutant DRE box.When fused to the GAL4 DNA-binding domain, either full-length QRAP2 or its N-terminus functioned effectively as atrans-activator in the yeast one-hybrid assay with its C-ter-minus completely inactive. Our data indicate that QRAP2could be a new member of the AP2/EREBP transcriptionfactor family involved in activation of down-stream targetgenes in response to environmental stress, especially underdrought conditions. 展开更多
关键词 分子无性繁殖 压条繁殖 DNA 遗传表达 活性激化 RAP2
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