两种根癌土壤杆菌介导的荧光素酶在植物体内表达分析  

Analysis of the Expression of Luciferase Mediated by Two Stains of Agrobacterium tumefaciens in Plants

作　　者：薛杨[1] 赵文军[2] 孙现超[1,3] 青玲[3] 杨水英[3] 

机构地区：[1]中国农业科学院柑桔研究所,重庆400712 [2]中国检验检疫科学研究院动植物检疫研究所,北京100029 [3]西南大学植物保护学院,重庆400715

出　　处：《西南大学学报（自然科学版）》2010年第12期63-66,共4页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(30900937);中国博士后科学基金资助项目(20090450798);中央高校基本科研业务费专项资金资助项目(XDJK2009B023)

摘　　要：根据报道的荧光素酶(Luciferase,Luc)基因设计引物,PCR扩增获得Luc基因,经BamH I和PstI酶切连接到载体pCHF3上,成功构建植物表达载体pCHF3-Luc.将pCHF3-Luc分别转入根癌土壤杆菌株EHA105和LBA4404细胞内,利用根癌土壤杆菌渗入法导入本氏烟和大麦,荧光检测仪内检测本氏烟和大麦叶片中瞬时表达的Luc活性.结果表明:两种根癌土壤杆菌介导的Luc可以在本氏烟中成功表达,而在大麦中不能表达;利用根癌土壤杆菌渗入法在本氏烟中表达外源基因时,EHA105菌株优于LBA4404菌株.Luciferase gene was amplified by PCR with primer pairs designed based on the reported paper.Having been digested with BamH I and Pst I,it was cloned into vector pCHF3 to construct plant expression vector pCHF3-Luc.pCHF3-Luc was then transformed into Agrobacterium strains EHA105 and LBA4404.The cells of Agrobacterium EHA105 and LBA4404 containing pCHF3-Luc were inoculated to barley （Hordeum vulgare L.） and Nicotiana benthmiana by agroinfiltration.The activities of Luc that were transiently expressed in the leaves of barley and N.benthmiana were detected with a luminometer.The results showed that Luc could be expressed in the leaves of N.benthmiana,but not in those of barley.EHA105 was better than LBA4404 when they were used to mediate the expression of foreign genes in N.benthmiana leaves.

关 键 词：根癌土壤杆菌 荧光素酶 表达 分析 

分 类 号：Q933[生物学—微生物学]