摘要
为了研究花粉致死基因ZmAA1的功能,进而创制转ZmAA1基因玉米不育新材料。在Gen Bank中找到花粉致死基因ZmAA1及启动子Pg47,并在目的片段的上下游设计増加酶切位点,基因合成构建到克隆载体puc57上,采用传统酶切连接的方法构建了植物表达载体pCAMBIA3300-Pg47-ZmAA1-35S-bar,并利用农杆菌介导法转化优良玉米自交系郑58萌动胚。成功构建植物表达载体pCAMBIA3300-Pg47-ZmAA1-35S-bar;共获得94株除草剂抗性植株,其中47株目的片段PCR呈阳性反应,对温室中表现为花粉败育的PCR阳性植株进行目的片段及筛选标记bar基因RT-PCR与蛋白免疫学试纸条检测,结果表明,花粉致死基因ZmAA1及启动子Pg47已经整合到玉米基因组并表达。
In order to study the function of pollen lethal gene ZmAA1 and create tansgenic male sterile materials. ZmAA1 and its specific promoter Pg47 were found according to Gen Bank,designed to plus restriction endonuclease on the downstream and inserted them into an cloning vector puc57,construction of vector pCAMBIA3300-Pg47-ZmAA1-35S-bar used traditional construction methods digested connection. A maize inbred line Zheng 58 was infected by Agrobacterium tumefacfaciens with the vector pCAMBIA3300-Pg47-ZmAA1-35S-bar,94 PPT-resistant seedlings were obtained and 47 plants were PCR positive. The pollen sterility in greenhouse RT-PCR assay for gene bar in the transgenic plant leaves and immuno strip test suggested that pollen lethal gene ZmAA1 had been integrated into the maize genome,and protein was expressed.
作者
孟令聪
宋广树
吕庆雪
刘宏伟
张志军
李春雷
王敏
刘文国
MENG Lingcong;LU Qingxue;ZHANG Zhijun;LI Chunlei;WANG Min;SONG Guangshu;LIU Hongwei;LIU Wenguo(Jilin Jinong Agricultural New and Hi-Tech Develop Co. , Ltd , Gongzhuling 136100 , China;Jilin Academy of Agricultural Sciences, Changchun 130124 , China)
出处
《华北农学报》
CSCD
北大核心
2016年第3期101-106,共6页
Acta Agriculturae Boreali-Sinica
基金
吉林省科技厅产业技术创新战略联盟项目(20140309005NY)
国家支撑计划项目(204BAD01B01)
