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  

Construction of the pCHF3-MAP65-1-YFP and the pCHF3-MAP65-2-YFP vectors with ligation-independent cloning method

作　　者：王培培 韩榕 WANG Pei-pei;HAN Rong(College of Life Science, Shanxi Normal University, Linfen 041000, China;Higher Education Key Laboratory of Plant Molecular and Environmental Stress Response, Linfen 041000,China)

机构地区：[1]山西师范大学生命科学学院,山西临汾041000 [2]植物分子与环境胁迫响应山西省高等学校重点实验室,山西临汾041000

出　　处：《江苏农业学报》2018年第2期266-270,共5页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(30671061);山西省自然科学基金项目(2008011059-1)

摘　　要：利用不依赖于连接反应的克隆(Ligation-independent cloning,LIC)方法构建了p CHF3-MAP65-1-YFP和p CHF3-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-l-YFP and pCHF3-MAP65-2-YFP vector that would be used for subsequent research of localization of the Arabidopsis thaliana microtubule binding protein MAP65-1and MAP65-2in the cell and the production mechanism of mitotic anomaly in UV-B radiation.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.

关 键 词：LIC方法 拟南芥 MAP65-1 MAP65-2 

分 类 号：S188[农业科学—农业基础科学]