抗草甘膦转基因大豆PCR检测及问题探讨

转基因植物的检测具有重要的意义。用抗草甘膦转基因大豆中的外源CaMV35S启动子、CP4 EPSPS和巢式PCR引物,应用PCR方法,从中扩增出预期大小的DNA片段。将扩增产物回收后测序,经同源性分析扩增产物为CaMV35S启动子和CP4 EPSPS的一部分序列。与常规PCR相比,巢式PCR在检测转基因大豆中具有更高的特异性。讨论了PCR检测过程中假阴性和假阳性的原因。

PCR Detection of Transgenetic Soybean Resistant to Glyphosate and Problem Discussion

It is important to identify the transgenetic plant.The anticipated fragments were amplified from DNA of transgenetic plant resistant to Glyphosate by PCR method using the primers of CaMV35S promoters,CP4-EPSPS and nested PCR.The fragments were purified from the Agrose gel and sequenced The homology analysis proved that the fragments were parts of CaMV35S promoters and CP4-EPSPS.Nested PCR was more accurate in identification of transgenetic soybean than common PCR.The reasons of false negative and positive results were also discussed.