以实时荧光定量PCR技术检测转基因玉米MON88017  

Detection of Genetically Modified Maize MON88017 by Quantitative Real- time PCR

作　　者：袁磊[1,2] 孙红炜[1] 李凡[1] 李宁[1,3] 赵蕾[3] 路兴波[1] 

机构地区：[1]山东省农业科学院植物保护研究所/山东省植物病毒学重点实验室,山东济南250100 [2]山东商务职业学院食品工程系,山东烟台264670 [3]山东师范大学生命科学学院,山东济南250014

出　　处：《作物学报》2011年第11期2117-2121,共5页Acta Agronomica Sinica

基　　金：国家转基因生物新品种培育科技重大专项(2008ZX08012-001);山东省农业科学院博士科研启动基金项目资助

摘　　要：根据转基因玉米MON88017的左侧侧翼序列和玉米内标基因（zSSIIb）分别设计特异性引物及Taqman探针,通过实时荧光PCR技术建立MON88017的定量特异性检测方法。利用含有内标基因和侧翼序列的标准质粒分子,建立了玉米内标基因和侧翼序列的标准曲线。检测了5个已知MON88017含量（0.01%、0.05%、0.10%、0.50%和1.00%）的混合样品中的转基因含量,结果表明检测底限可以达到19~30个阳性拷贝。该研究建立的实时荧光定量PCR技术灵敏度高、特异性强。Transgenic event-specific primers and Taqman probes based on the left-flanking sequence of MON88017 and endogenous gene （zSSIIb） were designed, and a real-time PCR method was developed to quantitatively detect the event-specific genetically modified maize. The reference molecule composed of endogenous gene and flanking sequence of MON88017 was constructed artificially. Two standard curves of the reference gene sequence and the 5’ flanking sequence were established. We detected five mixed samples, and their genetically modified contents of MON88017 were 0.01%, 0.05%, 0.1%, 0.5%, and 1%, respectively. The lowest detection limit was 19-30 copies. This study indicated that the Taqman probes real-time PCR is highly specific and sensitive.

关 键 词：转基因生物 实时荧光定量PCR MON88017 标准分子 

分 类 号：S513[农业科学—作物学]