利用不依赖连接反应的克隆方法构建pCHF3-MAP65-1-YFP和pCHF3-MAP65-2-YFP表达载体

摘要

利用不依赖于连接反应的克隆(Ligation-independent cloning,LIC)方法构建了pCHF3-MAP65-1-YFP和pCHF3-MAP65-2-YFP载体,用于研究拟南芥微管结合蛋白MAP65-1和MAP65-2在细胞中的定位及UV-B辐射下有丝分裂异常现象的产生机制.试验结果表明,相对于传统载体构建方法,LIC方法试验设计操作简便,耗时短,转化率高,可减少后期重组子筛选工作.%The method of ligation-independent cloning (LIC) was used to construct pCHF3-MAP65-1-YFP and pCHF3-MAP65-2-YFP vector that would be used for subsequent research of localization of the Arabidopsis thaliana microtu-bule binding protein MAP65-1 and MAP65-2 in the cell and the production mechanism of mitotic anomaly in UV-B radia-tion. The results showed that the method of LIC had simple operation,short time consuming,high conversion rate,and less workload of later screening recombinant,compared with the traditional method of building the carrier.