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| 转基因玉米特异性检测阳性标准分子的构建与应用 | |
| 引用本文： | 沈雨萌,李飞武,李葱葱,张明,陆军.转基因玉米特异性检测阳性标准分子的构建与应用[J].玉米科学,2010,18(4):52-57. |
| 作者姓名： | 沈雨萌 李飞武 李葱葱 张明 陆军 |
| 作者单位： | 1. 东北师范大学生命科学学院,长春,130021;吉林省农业学科院生物技术研究中心,长春,130033 2. 吉林省农业学科院生物技术研究中心,长春,130033 3. 东北师范大学生命科学学院,长春,130021 |
| 基金项目： | 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。 |
| 关 键 词： | 转基因玉米 转化体特异性检测 阳性标准分子 |
| 收稿时间： | 2009/3/30 0:00:00 |
Construction and Application of the Positive Standard Reference Molecule for Event-Specific Detection in Genetically Modified Maize |
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| SHEN Yu-meng,LI Fei-wu,LI Cong-cong,ZHANG Ming,LU Jun.Construction and Application of the Positive Standard Reference Molecule for Event-Specific Detection in Genetically Modified Maize[J].Journal of Maize Sciences,2010,18(4):52-57. | |
| Authors: | SHEN Yu-meng LI Fei-wu LI Cong-cong ZHANG Ming LU Jun |
| Abstract: | 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-specific products of Bt11 and MON810 maize were amplified. The pMD-zSSIIb and two PCR products were digested with the corresponding enzymes respectively, Bt11 and MON810 products were cloned into pMD-zSSIIb, obtained positive standard molecular pMD-ZB and pMD-ZM, which were confirmed by specific testing. The results showed that the obtained positive standard molecular could be used in the detection of genetically modified products as the positive control. |
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