马铃薯prp1-1启动子的克隆及转基因研究  

Cloning of the promoter of prp 1-1 gene from potato leaf and study on transgenic tobacoo

作　　者：马志刚[1] 鄢波[1] 王玲仙[1] 黄兴奇[1] 万萌[1] 郑丽屏 

机构地区：[1]云南省农业科学院生物技术研究所,云南昆明650223

出　　处：《西南农业学报》2002年第1期12-14,共3页Southwest China Journal of Agricultural Sciences

基　　金：云南省应用基础基金重点项目 (2 0 0 1-0 4-2 0 )

摘　　要：采用PCR技术 ,从云南马铃薯叶片DNA中分离克隆到了马铃薯病程相关蛋白 (prpl - 1)基因启动子 ,与国外文献报道序列完全一致。将这一启动子亚克隆到中间载体pUC18中 ,再构建到植物表达载体 pBI12 1上 ,以取代其原有的CaMV35S启动子 ,得到重组植物表达载体 pBI12 1M ;用电脉冲法将pBI12 1M转入农杆菌LBA4 40 4。应用农杆菌介导法 ,将携带有 pBI12 1M重组质粒的农杆菌转化烟草 ,经卡那霉素筛选 ,得到了一批转基因植株。转基因植株的PCR鉴定表明 ,启动子整合到了烟草植株基因组中。本研究为下一步通过X -Glu染色法检测启动子下GUS基因的表达情况 ,研究该启动子的病原菌特异性诱导活性 。A promoter of potato pathongensis related gene( prp 1 1)was cloned from the potato leaf DNA by using PCR.It was found that the nucleotide sequence of the promoter was utterly same as what reported in foreign references.The promoter was firstly subcloned into the intermediate vector pUC18,and then constructed into plant expression vector pBI121,which substituted for the CaMV35S promoter,thus a recombinated plant expression vector pBI121 obtained.By electroporation,pBI121M was transformed into Agrobacterium LBA4404.The Agrobacterium carring recombinant plasmid pBI121M was used to transform tobacco.Some transgenic tobacco plants were obtained through selection of Kanamycin.The PCR detection indicated that the promoter was integrated into the tobacco genome.By analysis of GUS gene expression,the selective induced activity of the promoter will be determined.

关 键 词：基因克隆 转基因 烟草 启动子 马铃薯 prp1-1 植株 病原性 蛋白基因 电脉冲法 

分 类 号：S336[农业科学—作物遗传育种] S532.037[农业科学—农艺学]