摘要
在拟南芥和水稻中Argonaute(AGO)蛋白是RNA介导的沉默复合体(RISC)的核心组分,在植物叶极性的分化方面具有重要的调节作用。实验根据AGO1基因序列设计一对特异引物,提取野生型拟南芥RNA作为模板,采用反转录PCR方法扩增出AGO1基因,并插入到克隆载体pGEM-T中。筛选鉴定后将AGO1连接到植物表达载体pBI121上,构建起植物表达载体pBI121-AGO1。并且利用农杆菌介导的方法转化拟南芥,通过抗性筛选,获得转基因植株。然后对转基因植株进行表型分析及AGO1蛋白的RT-PCR检测。通过对转基因拟南芥表达分析发现,与野生型拟南芥相比超表达AGO1蛋白的转基因拟南芥叶片明显呈锯齿状,说明AGO1基因影响拟南芥叶片发育。
In Arabidopsis thanalia and Oryza sativa ,Argonaute （AGO） proteins are the core components of RISC which play important roles in miRNA function, and AGO1 encodes an RNA Slicer that selectively recruits microRNAs and siRNAs. Total RNA was extracted from seedlings of Arabidopsis thanalia to clone the AGO1 gene. The plant expression vector of AGO1 was constructed （pBI121-AG01） and transformed into Arabidopsis by the Agrobacteriurn-mediated method. In our results, we se- lected the T1 transgenic Arabidopsis with AGO1 gene by Kanarnycin resistance and RT-PCR analysis. According to the pheno- type of transgenie plants, we discovered the incised margin of the transgenic Arabidopsis leaves.
出处
《河北科技大学学报》
CAS
2014年第1期51-57,共7页
Journal of Hebei University of Science and Technology
基金
山东省自然科学基金(ZR2012CQ021)
山东省高等学校科技计划项目(J12LE07)
