摘要
[目的]研究MYB类基因At3g11280和At5g05790的详尽功能。[方法]利用农杆菌将标签蛋白融合的At3g11280和At5g05790基因转化拟南芥,利用转基因植株中标签蛋白的抗体进行染色体免疫共沉淀(ChIP)试验,挖掘两基因所调控的下游基因。[结果]首先扩增了At3g11280和At5g05790基因的CDS片段,随后经过一系列的亚克隆过程将两个基因和标签蛋白的融合基因片段克隆到载体pWM101上,位于35S启动子的下游,转录受35S启动子的调控。[结论]成功地构建了标签融合蛋白植物表达载体,两个基因分别携带HA和Flag标签,载体的成功构建将为ChIP试验奠定基础。
[Objective] The aim was to study the detailed function of MYB genes of At3g11280 and At5g05790.[Method] The Agrobacterium was used to transform the tag-protein-fused At3g11280 and At5g05790 genes into Arabidopsis and the antibody of the tag protein in the transgenic plants was used to conduct the chromosomes immune precipitation（ChIP） experiment so as to excavate downstream genes controlled by above 2 genes.[Result] Firstly,the CDS fragments of At3g11280 and At5g05790 genes were amplified,and then through a series of the subcloning process,the 2 genes and the fused gene fragments and tag protein could be cloned into the pWM101 carrier,which located in the downstream of 35S promoter and its transcription was controlled by 35S promoter.[Conclusion] The plant expression vector with tag protein fusion was successfully constructed and 2 genes carried the tags of HA and Flag respectively.The successful construction of the vector will lay the foundation for ChIP test.
出处
《安徽农业科学》
CAS
北大核心
2011年第34期20949-20950,21006,共3页
Journal of Anhui Agricultural Sciences
