| 花粉致死基因ZmAA1对玉米遗传转化的研究 |
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| 引用本文： | 孟令聪,宋广树,吕庆雪,刘宏伟,张志军,李春雷,王敏,刘文国.花粉致死基因ZmAA1对玉米遗传转化的研究[J].华北农学报,2016(3):101-106. |
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| 作者姓名： | 孟令聪 宋广树 吕庆雪 刘宏伟 张志军 李春雷 王敏 刘文国 |
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| 作者单位： | 1. 吉林吉农高新技术发展股份有限公司,吉林 公主岭,136100;2. 吉林吉农高新技术发展股份有限公司，吉林 公主岭 136100; 吉林省农业科学院，吉林 长春 130124 |
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| 基金项目： | 吉林省科技厅产业技术创新战略联盟项目(20140309005 NY)；国家支撑计划项目(204BAD01 B01) |
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| 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。
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| 关 键 词： | 玉米 ZmAA1 基因 启动子 Pg47 遗传转化 |
Study on Transformation in Maize of Pollen Lethal Gene ZmAA1 |
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| Abstract: | In order to study the function of pollen lethal gene ZmAA1 and create tansgenic male sterile materi-als.ZmAA1 and its specific promoter Pg47 were found according to GenBank,designed to plus restriction endonu-clease 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 integrat-ed into the maize genome,and protein was expressed. |
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| Keywords: | Maize ZmAA1 gene Pg47 promoter Genetic transformation |
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