LAMP在检测转基因抗草甘膦大豆cp4-epsps基因上的应用

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。

Application of LAMP in cp4-epsps Gene Detection of Roundup Ready Soybean

Taking glyphosate-tolerant soybean as the research object,loop-mediated isothermal amplification(LAMP) was used.Four specific primers were designed according to the six regions of 5-enolpyruvlshimimate-3-phosphate synthase.The amplification could be obtained in 30 min by incubating all of the reagents in a single tube with Bst DNA polymerase at 65 °C.Results showed that sensitivity of the LAMP assay were 10 times higher than conventional PCR assay on the detection limit for cp4-epsps gene.Results clearly demonstrated that the LAMP-based assay was a sensitive and reliable means for the detection of glyphosate-tolerant soybean.