荣晶,阮孟斌,王斌,彭明.木薯MeMYB63 基因特性及启动子活性初步分析[J].广东农业科学,2013,40(7):129-133 |
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木薯MeMYB63 基因特性及启动子活性初步分析 |
Preliminary analysis on genetic characteristics and the promoter activity of MeMYB63 in cassava (Manihot esculenta Crantz) |
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DOI: |
中文关键词: MeMYB63 基因 转录因子 蛋白亚细胞定位 转录激活 |
英文关键词: MeMYB63 gene transcription factor protein subcellular location transcriptional activation |
基金项目:国家重点基础研究发展规划(2010CB126602);海南省重大科技专项(ZDEX2013023-1-10);海南省自然科学基金(312049) |
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中文摘要: |
为了解木薯干旱相关的MeMYB63 基因的基本特性,从木薯中克隆了干旱诱导的调控基因MeMYB63的全长cDNA 及起始密码子上游1 500 bp 的DNA 片段,在转基因拟南芥中对该基因的启动子活性进行了初步分析。利用绿色荧光蛋白(green fluorescent protein,GFP)对MeMYB63 蛋白进行标记,通过烟草瞬时转化系统观察融合蛋白的亚细胞定位。利用酵母转化试验确认MeMYB63 是否具有转录激活功能。结果表明,MeMYB63 基因5' 上游1543 bp 的片段具有明显启动子特征。MeMYB63 蛋白的亚细胞定位试验发现MeMYB63 与GFP 融合的蛋白产物仅在细胞核出现,酵母转录激活试验表明,MeMYB63 具有明显的转录激活功能,说明该基因具有转录激活结构域。亚细胞定位及转录激活试验结果表明,MeMYB63 可能是一个转录因子,为今后进行该基因功能的研究提供了重要依据。 |
英文摘要: |
In order to understand the basic characteristics of cassava MeMYB63 gene which relates to drought stress,we cloned its full length cDNA and the 1 500 bp DNA fragment upstream of MeMYB63 gene initiation codon, and preliminarily analyzed the gene promoter activity in transgenic Arabidopsis plants. By using the green fluorescent protein (GFP) MeMYB63 protein was labeled , subcellular localization was operated by means of tobacco transient transformation system to observed fusion protein. And the transcriptional activation function of MeMYB63 protein was confirmed in the yeast transformation experiment. The results showed that the typical promoter elements were found in the 5 ' upstream 1543 bp DNA fragment. MeMYB63 and GFP fusion protein just existed in the nucleus during the subcellular localization experiment of the MeMYB63 protein. Moreover, the result of yeast transcriptional activation experiment showed that MeMYB63 had obvious transcriptional activation function with a transcription activation domain. According to all of the results of the subcellular localization and transcriptional activation, MeMYB63 might be considered as a transcription factor, which provided an important basis for the future study of the gene function. |
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