TaqMan探针用于转基因食品的荧光定量PCR检测

选择了内源基因大豆植物凝集素 (lectin)、玉米转化酶 (invertase)和外源基因抗草甘膦除草剂的 5 烯醇式丙酮酰莽草酸 3 磷酸合成酶 (cp4epsps)、抗欧洲玉米螟的苏云金芽孢杆菌杀虫毒蛋白 (cryIAb)的TaqMan探针 ,确定了探针浓度和镁离子浓度等反应条件 ,分别对转基因大豆和转基因玉米系列标准品进行内源基因和外源基因的荧光PCR扩增 ,在PCR反应过程中分别以TET和FAM两种荧光通道信号分别追踪同一样品DNA的内源基因和外源基因的扩增动力学变化 ,并依此绘制了ΔCt值与转基因食品百分比含量之间的标准曲线 ,建立了转基因大豆和转基因玉米的TaqMan -荧光定量PCR检测方法 ,该方法具有探针设计较简便、成本较低的特点 ,初步实现了对转基因食品的定量分析及品种鉴定。

Fluorescence Quantitative PCR Detection of Genetically Modified Food by TaqMan Probe

Primers and TaqMan probes of lectin, invertase, cp4epsps, and cryIAb were designed The reaction conditions such as probe concentration and MgCl 2 concentration were optimized GM soybean and GM maize reference materials were detected by fluorescence PCR amplification of endogenous gene and exogenous gene The standard ΔCt curves of endogenous gene and exogenous gene vs GM content in reference materials were generated and a linear regression equation was obtained TaqMan fluorescence quantitative PCR method was established to detect GM soybean and GM maize The design of probe features simplicity and low cost. The method has been used for quantitative analysis and variety identification of genetically modified food