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以转基因抗草甘膦大豆为主要研究对象,利用环介导等温扩增技术(Loop-Mediated Isothermal Amplification,LAMP),针对cp4-epsps合成酶基因(5-enolpyruvlshimimate-3-phosphate synthase)的6个区域设计4条特异性引物,利用一种链置换DNA聚合酶(BstDNA polymerase),在65℃保温30 min,通过荧光显色即可完成对转基因的检测工作。结果显示,该LAMP方法能够特异性检测cp4-epsps基因,其检测灵敏度是常规定性PCR方法的10倍。建立了针对转基因大豆cp4-epsps基因的LAMP检测方法,其具有高度的特异性及稳定性,结果可靠,适合转基因抗草甘膦大豆的快速检测。
Glyphosate-tolerant soybeans were selected as the main research object. Six loops of 5-enolpyruvlshimimate-3-phosphate synthase (cp4-epsps) gene were amplified by loop-mediated Isothermal Amplification (LAMP) Four specific primers were designed in the region to detect the transgenes by fluorescence colorization using a strand displacement DNA polymerase (BstDNA polymerase) at 65 ° C for 30 min. The results showed that the LAMP method can detect the cp4-epsps gene specifically, and its detection sensitivity is ten times that of the conventional qualitative PCR. A LAMP assay for gene cp4-epsps in transgenic soybean was established, which has high specificity and stability and reliable results, suitable for the rapid detection of transgenic glyphosate-resistant soybean.