摘要
以转基因抗虫和耐除草剂玉米Bt11为研究材料,建立Bt11环状等温扩增快速检测技术方法。利用一种具有链置换活性的BstDNA聚合酶,针对Bt11中玉米基因组与外源基因结合处序列,设计4条特异引物,并对反应温度、时间、灵敏度、结果判断方法等参数进行初步探索。结果显示,LAMP检测方法最适反应温度及反应时间分别为65℃和60min,其灵敏度为常规定性PCR的20倍。LAMP技术检测转基因玉米品系Bt11具有特异性高、快速、简便等优点,具有广阔的应用前景。
A rapid assay method for insect-resistant and herbicide-tolerant maize Bt11 was developed based on the loop-mediated isothermal amplification （LAMP） . The event-specific sequence was amplified by the Bst DNA polymerase and four primers which were designed according to the six regions of the unique and specific integration junction sequences between the host plant genome DNA and the integrated gene of Bt11. The reaction time,temperature, sensitivity,result-determining were optimized. The results showed that the optimum temperature and time was 63℃ and 60 min,respectively. The sensitivity of LAMP detection was 0. 0005% ,which was 20 times more sensitive than the conventional PCR. Results indicated that LAMP can provide a sensitive,specific,stable and reliable test for the detection of GM maize Bt11.
出处
《华北农学报》
CSCD
北大核心
2010年第4期49-52,共4页
Acta Agriculturae Boreali-Sinica
基金
科技部国际合作项目(2006DFA32380)
天津市农业科学院院长基金项目(07013)
关键词
转基因玉米
环状等温扩增
检测
GM maize
Loop-mediated isothermal amplification （LAMP）
Detection
