影响拟南芥转化效率和激活标签丢失的因素分析  

Analysis of factors affecting transformation efficiency and loss of enhancer in generation of activation-tagged mutants of Arabidopsis thaliana

作　　者：王爱荣[1] 吴智芳[1] 张丽丽[1] 张春华[1] 张冬梅[1] 鲁国东[1] 周洁[1] 王宗华[1] 

机构地区：[1]福建农林大学教育部生物农药与化学生物学重点实验室福建农林大学功能基因组学中心,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2006年第3期298-302,共5页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：国家自然科学基金(30471178);福建省自然科学基金(B0210018)资助项目

摘　　要：用激活标签载体pSK I015和农杆菌介导的浸花转化法创建拟南芥激活标签突变体,优化了浸花法转化方案,采取原地转化的措施,减少了工作量,又保证了植株在转化期间的正常生长,转化效率高达1.434%.突变体分析表明,57.5%含单拷贝T-DNA插入,100%整合了Bar基因,87%含有CaMV 35S增强子,进一步分析发现CaMV 35S增强子丢失机率与农杆菌继代次数呈正相关.To increase transformation efficiency, we modified the procedure of floral dipping method, and transformed plants at the original site during the generation of acitation tagging mutants library of Arabidopsis thaliana. This modification reduced workload, guaranteed the normal growth of plants and achieved relatively high transformation efficiency up to 1.434%. The mutant library was analyzed by detecting T-DNA copy number or the enhancer genes and Bar gene. The result showed that 57.5% mutants contained single-copy T-DNA, 100% harbored Bar gene and 87% were activation-tagged. Further analysis showed that the rate of loss of CaMV 35S enhancers was positively correlated to the times of culturing Agrobacterium tumefaciens.

关 键 词：拟南芥 农杆菌 激活标签 转化 

分 类 号：Q94-331[生物学—植物学]