抗草甘膦转基因大豆PCR检测及问题探讨

PCR Detection of Transgenetic Soybean Resistant to Glyphosate and Problem Discussion

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。

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.