拟南芥CBF1基因植物表达载体构建及其对野生蕉的遗传转化  

Construction of plant expression vector containing CBF1 and its genetic transformation in wild banana

作　　者：刘凯[1,2] 胡春华[1] 魏岳荣[1] 易干军[1] 邵秀红[1] 

机构地区：[1]广东省农业科学院果树研究所,广东广州510640 [2]湖南农业大学园艺园林学院,湖南长沙410128

出　　处：《湖南农业大学学报（自然科学版）》2011年第3期248-252,共5页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家自然科学基金项目(31000903);广东省自然科学基金项目(10451064001006326);广东省农业科学院院长基金项目(20090104)

摘　　要：从拟南芥中克隆CBF1基因,转入到植物表达载体pBI121的XbaI和SacI位点,得到中间重组质粒pBI121-CBF1,用EcoRI和HindⅢ将35S启动子与CBF1基因从pBI121-CBF1切下,转入到植物表达载体pCAMBIA1301中,构建植物表达载体p1301-CBF1。将构建的植物表达载体转入农杆菌EHA105,采用农杆菌介导法转化野生蕉胚性悬浮细胞,经GUS组织化学染色和PCR鉴定,证明CBF1基因已转入野生蕉中。The CBF1 gene（CRT/DRE binding factor 1） was obtained from Arabidops＆ thaliana by PCR and cloned into pMD 18-T. Then CBF1 gene was sub-cloned into pBI 121 vector to abtain the middle recombinant plasmid pB1121-CBF 1 via Xba I and Sac I restriction enzyme sites. After that, the fragments containing 35S-CBF1 were digested from recombinant plasmid pBI121-CBF1 by EcoR I and Hind Ⅲ, and directionally linked to the plant expression vector pCAMBIA1301 to get the plant expression vector p1301-CBF1. We transformed the embryogenic cell suspensions of wild banana（Musa intinerans Cheesm） with pl301-CBF1 via Agrobacterium-mediated transformation. Results from histochemical GUS assay and PCR suggested that CBF1 gene has been transferred into the wild banana.

关 键 词：拟南芥 CBF1基因 野生蕉 遗传转化 胚性悬浮细胞 

分 类 号：Q785[生物学—分子生物学]