转基因玉米特异性检测阳性标准分子的构建与应用  

Construction and Application of the Positive Standard Reference Molecule for Event-Specific Detection in Genetically Modified Maize

作　　者：沈雨萌[1,2] 李飞武[2] 李葱葱[2] 张明[2] 陆军[1] 

机构地区：[1]东北师范大学生命科学学院,长春130021 [2]吉林省农业学科院生物技术研究中心,长春130033

出　　处：《玉米科学》2010年第4期52-57,共6页Journal of Maize Sciences

基　　金：吉林省财政厅科研育种专项(2007)

摘　　要：试验用PCR方法从玉米中扩增内源基因zSSIIb片段,并将其克隆到pMD18-T载体上,获得中间载体pMD-zSSIIb;根据Bt11和MON810玉米转化体特异性序列,分别设计带酶切位点的引物,扩增出Bt11和MON810转化体特异性产物;用相应的酶对pMD-zSSIIb和两种PCR产物进行酶切,分别将Bt11和MON810产物克隆到pMD-zSSIIb上,获得阳性标准分子pMD-ZB和pMD-ZM,并对其进行特异性测试。结果表明,获得的阳性标准分子可以作为转基因产品检测时的阳性对照。The positive standard reference molecule for event-specific detection of Bt11 and Mon810 maize were established.The maize endogenous gene zSSIIb was amplified using the PCR method,then cloned into pMD18-T plasmid vector and obtained the intermediate recombinant plasmid pMD-zSSIIb.The primers with different enzyme sites were designed according to the 3 -flanking sequences of Bt11 and MON810 maize,respectively,and event-spe-cific products of Bt11 and MON810 maize were amplified.The pMD-zSSIIb and two PCR prod...

关 键 词：转基因玉米 转化体特异性检测 阳性标准分子 

分 类 号：S51[农业科学—作物学]