转Bt基因水稻种子中Bt蛋白含量测定方法的研究

采用ELISA技术检测转Bt基因水稻中Bt蛋白的含量,判断水稻样品中是否含有转基因成分。利用研磨、酶标、孵育等技术对样品进行前处理。采用阳性质控物浓度等倍稀释方法,建立标准曲线,相关系数为0.997 4。用一系列不同转基因含量的标准基体材料,分析方法的最低检测限,灵敏度达0.1%。通过对我国进入生产性试验的转Bt基因水稻品系TT51-1和科丰6号的测试,表明该方法与普通PCR方法真实性和灵敏度一致,可以广泛应用于转Bt基因水稻及其粗加工产品的转基因成分检测。为转基因生物安全监管和安全性评价提供技术支撑。

Protein Content in Bt-Gene Rice Seed

In this study,the enzyme-linked immunosorbent assay(ELISA) has been develpoed for the detection of Bt protein content in transgenic Bt rice to evaluate the presence of genetically modified ingredients in rice samples.The sample pre-treatment step involved in grinding,enzymatic labeling reaction and incubation process.The standard curve was constructed by dilution of the the positive control material with the same dilution factor,and the correlation coefficient is 0.997 4.A range of different standards containing of genetically modified ingredients were used to analyze the limit of detection method.The results showed the detection sensitivity is up to 0.1%.Through testing the transgenic Bt rice strain TT51-1 and Kefeng 6 which are putting into the production test in China,the ELISA method developed demonstrated consistency with ordinary PCR methods in both authenticity and sensitivity aspects.Therefore,this ELISA method could be widely applied to detect genetically modified ingredients in transgenic Bt rice or raw commodities,providng technical support for the safety regulation and evalution of genetically modified organisms.