一种快速高效检测转基因玉米方法的建立  

Establishing of a Fast and Efficient Testing Method of Transgenic Maize

作　　者：周德龙 孟令聪[1] 郑淑波[1] 王楠[1] 李穆 王薪淇 卢实 王敏 刘文国[1] 路明[1] Zhou Delong;Meng Lingcong;Zheng Shubo;Wang Nan;Li Mu;Wang Xinqi;Lu Shi;Wang Min;Liu Wenguo;Lu Ming(Maize Research Institute,Jilin Academy of Agricultural Sciences/National Engineering Laboratory for Maize(Changchun)/National Engineering Research Center for Maize(Jilin)/Key Laboratory of Biology and Genetic Improvement of Maize in Northeast Region,Ministry of Agriculture and Rural Affairs/Open Laboratory of Jilin Province Crop Breeding of South Breeding Base,Changchun 130033,Jilin,China;Jilin Jinong Hi-Tech Development Co.,Ltd.,Gongzhuling 136100,Jilin,China)

机构地区：[1]吉林省农业科学院玉米研究所/玉米国家工程实验室(长春)/国家玉米工程技术研究中心(吉林)/农业农村部东北中部玉米生物学与遗传育种重点实验室/吉林省农作物育种南繁基地开放实验室,130033,吉林长春 [2]吉林吉农高新技术发展股份有限公司,136100,吉林公主岭

出　　处：《作物杂志》2022年第1期65-69,共5页Crops

基　　金：国家转基因重大专项养分高效利用转基因玉米新品种培育(2016ZX08003-005);国家科技支撑计划东北中北部玉米商业化育种技术研究与示范(2014BAD01B01)。

摘　　要：建立高效的目的基因检测方法是保障生物育种研发和产业化进程的重要技术支撑。应用常规PCR法、TaqMan探针实时荧光PCR法及叶片直接PCR法对转基因玉米叶片进行CaMV35S启动子和NOS终止子2个转基因元件的检测。结果表明,叶片直接PCR法分别较常规PCR方法和TaqMan探针实时荧光PCR法节约62%、48%的时间和25%、43%的成本,而且扩增出的目的条带清晰可见,符合目的片段大小,结果真实可靠,适用于大量转基因玉米植株叶片的快速筛选和鉴定。An efficient target gene detection method is an important technical support to ensure the development and industrialization of biological breeding.In this study,conventional PCR method,TaqMan probe real-time fluorescence PCR method and leaf direct PCR method were used to detect two transgenic elements of CaMV35S promoter and NOS terminator in leaves of transgenic maize plants.The experiment results showed that the leaf direct PCR method saved 62%and 48%time and reduced 25%and 43%cost compared with the conventional PCR method and TaqMan probe real-time fluorescence PCR method,respectively,and the amplified target band was clearly visible,and the fragment size was in accord with the target fragment size.The results were true and reliable,which was suitable for rapid screening and identification of a large number of transgenic maize leaves.

关 键 词：玉米 转基因 常规PCR法 TaqMan探针实时荧光PCR法 叶片直接PCR法 

分 类 号：S513[农业科学—作物学]