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酵母双杂交表达载体pGBKT7_NPR1与pGADT7_NPR1的构建及酵母菌的转化

Construction and Transformation of Yeast Two-hybrid Expression Vectors pGBKT7_NPR1 and pGADT7_NPR1
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摘要 NPR1是水杨酸介导的系统获得性抗性(SAR)途径中不可或缺的一个转录辅助因子。在SA诱导下,NPR1被转运至细胞核内并与一些转录因子结合,以启动下游基因的表达。本研究通过PCR扩增获得拟南芥中NPR1基因CDS序列,经过限制性内切酶酶切后在T4连接酶作用下分别克隆至pGBKT7与pGADT7酵母表达载体上,并成功将其转化至Gold酵母菌株中。为利用酵母双杂交筛选NPR1参与形成的转录复合体中其它可能存在的蛋白质并阐明其在植物防御信号传导途径中的互作模式奠定了基础。 NPR1 is an indispensable transcription co-factor in SAR induced by salicylic acid. Upon the SA stimuli, NPR1 is translocated into the nucleus and interacts with some transcription factors, and then starts the expression of down- stream genes. In this study, CDS sequence of NPR1 was cloned into two yeast expression vectors pGBKT7 and pGADT7, and then was transformed into yeast Gold strain. This study laid a foundation for sifting other possible proteins in transcrip- tional complex with participation of NPR1 by Yeast Two - Hybrid and elucidating its interaction model in plant defense sig-nal transduction pathways.
作者 肖牧 徐健 何乐 刘春林 阮颖
机构地区 湖南农业大学生物科学技术学院 湖南农业大学作物种质创新和资源利用国家重点实验室培育基地
出处 《作物研究》 2013年第3期213-216,共4页 Crop Research
基金 国家自然科学基金项目(31071455)
关键词 NPR1 基因克隆 酵母双杂交载体 酵母转化 NPR1 Gene cloning Yeast two - hybrid expression vectors Yeast transformation
分类号 Q78 [生物学—分子生物学]
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参考文献13

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