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作 者:韩素英[1] 张守攻[1] 汪泉[1] 王建华[1] 齐力旺[1]
出 处:《农业生物技术学报》2007年第3期446-450,共5页Journal of Agricultural Biotechnology
基 金:国家重点基础研究发展规划(973)(No.G19990160);国家自然科学基金项目(No.30571517);国家高技术研究发展计划(863)项目(No.2006AA100109);国家转基因与产业化专项(No.J2002-B-005;JY03-B-28-02)资助
摘 要:通过基因合成构建了脱水反应元件(dehydration-responsive element,DRE)酵母表达载体,通过同源重组,利用SD-His-Ura平皿筛选获得了含有pHis-1-DRE和pLacZi-DRE双重报告子的酵母即得DRE报告子。建立小叶杨(Populus simonii)干旱模型,提取RNA,构建小叶杨cDNA AD融合表达文库,通过酵母单杂交的方法,用DRE报告子对小叶杨cDNAAD融合表达文库进行了筛选,得到了2个阳性克隆。结果表明,所构建的DER报告子被激活,可用于小叶杨DER转录因子的克隆。The dehydration-responsive element (DRE) yeast expression vector was constructed by gene synthesis method and through the homologous recombination and screening the SD/-His/-Ura disc, and the DRE yeast reporter which containing the pHIS-1-DRE and pLacZi-DRE dual reporter yeast was obtained. Then the drought-resistance experimental model of Poplus simonii was established and the cDNA AD fusion expression library of P.simonii was constructed, the cDNA AD fusion expression library of P.simoni was screened by using the DRE reporter in the yeast one-hybrid System, in which two positive clones were obtained. The results indicated that the constructed DRE reporter could be activated and it could be used for the cloning of the DRE transcription factors from P.simonii.
关 键 词:酵母单杂交 DRE 顺式作用元件 转录因子 小叶杨
分 类 号:S188[农业科学—农业基础科学]
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