紫花苜蓿MsZFN基因超表达载体的构建及对烟草的转化  

Cloning of MsZFN Gene from Alfalfa and Transformation into Tobacco

作　　者：晁跃辉[1,2] 康俊梅[1] 王凭青[2] 杨青川[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193 [2]重庆大学生物工程学院,重庆400030

出　　处：《中国草地学报》2010年第6期21-26,共6页Chinese Journal of Grassland

基　　金：现代牧草产业技术体系岗位专家项目;国家重点基础研究发展计划(2007CB108905-2);国家科技支撑计划课题"中部温带地区优质高产;抗逆牧草新品种选育及产业化示范"(2008BADB3B05)

摘　　要：对紫花苜蓿一种新的CCCH类型锌指蛋白基因进行克隆及转化到烟草中,为研究该基因的功能提供依据。将紫花苜蓿锌指蛋白MsZFN基因(EU624138)连接到载体pBI121上,构建成植物表达载体pBI-ZFN。将表达载体转化到感受态农杆菌株LBA4404中,利用农杆菌介导的方法,以烟草无菌苗叶片为外植体,转化烟草,经卡那霉素筛选获得十几个烟草再生植株。对其中四个再生植株进行PCR和Southern检测,证明目的基因已经整合到烟草基因组中,通过RT-PCR检测,初步证明目的基因在烟草中可以表达。A new gene encoding a CCCH-Type zinc finger protein was cloned and transformed into tobacco to study its functions.An alfalfa zinc finger protein（MsZFN） gene（Access Number： EU624138） was obtained using RT-PCR technology,which was inserted into plasmid pBI121,and a plant expressing vector pBI-ZFN was constructed.Leaf segments of tobacco were inoculated by Agrobacterium tumefaciens LBA4404 with plasmid pBI-ZFN.PCR and PCR-Southern analysis of over 10 kanamycin resistant plants showed that the target gene had been integrated into the genomes of the tobacco.And expression of MsZFN gene had been confirmed in four tobacco plants by RT-PCR.

关 键 词：紫花苜蓿 锌指蛋白 烟草 转化 

分 类 号：S54[农业科学—作物学]