玉米ZmERD4基因的分离及转基因株系的获得  

Isolation of ZmERD4 Gene and Generation of Transgenic Maize Plants

作　　者：曹言勇[1] 赵心萍[1,2] 赵秋勇[2] 张艳[1] 唐保军[1] 刘方方[1] 王利锋[1] 李会勇[1] 

机构地区：[1]河南省农业科学院粮食作物研究所/河南省玉米生物学重点实验室,河南郑州450002 [2]太原理工大学化工学院,山西太原030024

出　　处：《河南农业科学》2011年第12期43-47,共5页Journal of Henan Agricultural Sciences

基　　金：国家转基因生物新品种培育重大专项(2009ZX08009-061B);河南省科技攻关项目(092102110039)

摘　　要：为了获得转ZmERD4基因玉米植株,验证ZmERD4在玉米育种中的应用价值,通过同源克隆的方法在抗旱性强的玉米自交系CN165中克隆了ZmERD4基因全长cDNA,该基因全长2 073bp,包含一个编码591个氨基酸的开放阅读框;推导的氨基酸序列和水稻中的OsERD4(XP476646)有82%的相似性,与拟南芥中的AtERD4(BAB63915)有58%的相似性。构建了由花椰菜花叶病毒35S启动子控制下的ZmERD4基因的植物表达载体,并利用农杆菌介导的方法对玉米杂交种Hi-Ⅱ进行了遗传转化,通过双丙氨磷抗性筛选、双丙氨磷抗性基因和ZmERD4基因PCR扩增对转基因后代株系进行了检测,获得了42个转ZmERD4基因株系,转化率为2%,为玉米抗旱育种提供了重要的物质基础。Using homology cloning method,the full length cDNA of ZmERD4 was isolated from inbred line CN165 of drought resistant maize.The length of ZmERD4 was 2073bp,and the predicated protein comprised of 591 amino acids.Bioinformatics analysis indicated that the amino acids shared 82% and 58% similarity with rice OsERD4（XP476646）and Arabidopsis AtE4RD4（BAB63915）,respectively.The plant transformation vector,in which ZmERD4 was driven by cauliflower mosaic virus 35S promoter,was conducted and transformed into the immature embryos of maize hybrid Hi-Ⅱ through Agrobaterium-mediated transformation system.Transgenic maize plants were assayed by bialaphos-resistance screening and PCR amplification of the bialaphos-resistance gene and ZmERD4 gene.The results showed that 42 transgenic maize lines with ZmERD4 were obtained,and the average transformation efficiency was 2%,which provided the important material basis for drought resistant maize breeding.

关 键 词：玉米 ZmERD4 植物转化载体 转基因株系 

分 类 号：S513[农业科学—作物学]