GA21转基因玉米实时荧光PCR检测方法的建立

成功建立了实时荧光PCR鉴定检测转基因玉米GA21品系的方法。该方法通过GA21玉米品系的OTPmEPSPS边界的270bp和133bp靶序列,设计品系特异性检测引物和探针,同时针对Pactin1mEPSPS边界的430bp靶序列设计品系特异性检测引物,应用实时荧光PCR和PCR技术,特异性检测GA21玉米品系。结果表明,应用实时荧光PCR的TaqMan探针技术检测转基因作物边界序列,不仅可以达到品系鉴定的目的,而且该方法和常规PCR比特异性强,简便快速,同时实验采用完全闭管检测,又降低了污染机会,为转基因作物的品系鉴定检测提供了新方法。

Establishment of the Real-time Fluorescent PCR Detection Method for Genetically Modified GA21 Maize

A event-specific real-time fluorescent PCR (RTF PCR) method was established for detection and identification of the genetically modified GA21 maize.The PCR primers and TaqMan probes were designed based on the GA21 border sequence of 270bp and 133bp of OTP/mEPSPS and 430bp of P actin 1/mEPSPS.The results show that the TaqMan probe could detect and identify GA21 maize specifically.RTF PCR method is much convenient and quicker to operate than common PCR,much less contamination will occur because whole detection process is finished in the contained tubes.RTF PCR could be a new method for detection of genetically modified organism event-specifically. ;