玉米隐性核不育保持系表达载体构建  

Expression Vector Construction of Recessive Genic Male Sterility Maintainer Line of Maize

作　　者：宋广树[1,2] 刘文国[1,2] 吕庆雪[1] 刘宏伟[1,2] 李春雷[1] 王敏[1] 王晶[1] 张志军[1,2] 

机构地区：[1]吉林吉农高新技术发展股份有限公司,吉林公主岭136100 [2]吉林省农业科学院,长春130124

出　　处：《玉米科学》2016年第2期35-39,46,共6页Journal of Maize Sciences

基　　金：吉林省科技厅产业技术创新战略联盟项目"新型雄性不育化玉米种子生产技术"(20140309005NY);国家支撑计划项目"春玉米区商业化育种技术研究与示范项目"(204BAD01B01)

摘　　要：在Genebank中搜索花粉致死基因Zm AA1、花粉育性恢复基因Ms45、粉质胚乳突变基因Mucronate及各基因特异调控因子和终止子序列,并在目的片段的上下游设计增加同尾酶Spe I(Nhe I、Xba I)和酶切后具有各种类型的DNA片段的Esp3I酶切位点,基因合成构建到克隆载体puc57上,命名为puc-Zm AA1、puc-Ms45、puc-Mc。采用传统酶切连接的方法将目的片段构建到植物表达载体p CAMBIA3300上,并用热击法将重组质粒导入农杆菌LBA4404及EHA105中,酶切、PCR检测及核苷酸序列测定证明,植物表达载体构建成功。According to Genbank, Zm AA1, Ms45, Mucronate（Mc） target gene and its specific promoter and ter-minator sequence were found, and design to plus restriction endonuclease on the downstream of fragment of isocau-domers Spe I（Nhe I, Xba I）, synthetic gene inserted into a cloning vector puc57, named puc-Zm AA1, puc-Ms45 andpuc-Mc. Target gene was inserted into an expression vector p CAMBIA3300 used traditional construction methodsdigested connection, and induced into Agrobacterium line LBA4404 and EHA105 by freeze-thaw method. Confirmplant expression vector was constructed successfully through digestion, PCR detection and homologous analysis.

关 键 词：玉米 Mucronate基因 Ms45基因 ZmAA1基因 表达载体 

分 类 号：S513.035.3[农业科学—作物学]