第一、二代转cp4-epsps基因大豆鉴定方法的建立

Establishment of detection method for the 1st and 2nd generation of cp4-epsps genetically modified soybeans

  • 摘要: 目的建立一种广谱性鉴定第一、二代转cp4-epsp基因大豆的检测方法,为准确鉴定第一、二代转cp4-epsps基因大豆及其产品提供技术支持.方法根据第一、二代转基因大豆的cp4-epsp基因保守序列设计一对能同时鉴定两代转基因大豆外源基因cp4-epsps的引物和探针,建立一种广谱性鉴定第一、二代转cp4-epsps基因大豆的方法,并从准确性、特异性、灵敏性及重复性4个方面对该方法进行评估.结果设计的引物/探针对鉴定第一、二代转cp4-epsps基因大豆及其产品具有广谱性;建立的检测方法能检测到反应体系中低至5×100拷贝的cp4-epsps基因,Ct为38.88,且能同时检测出两代转基因大豆.准确性和特异性评估结果显示仅两代转cp4-epsp基因的大豆能被检测到;40次重复试验结果显示,反应体系中5×100拷贝的cp4-epsps基因片段的检出率为100%.结论建立的第一、二代转cp4-epsps基因大豆鉴定方法具有特异强、灵敏度高、重复性好、准确性高等特点,可用于转cp4-epsp基因大豆及其产品的监测.

     

    Abstract: ObjectiveA widely adaptive detection method for the 1st and 2nd generation of cp4-epsps genetically modi-fied(GM) soybeans was established in order to provide technical support for accurately detecting the 1st and 2nd genera-tion of cp4-epsps GM soybeans and related products. MethodA pair of primer and probe was designed which was able to identify exogenous gene cp4-epsps of two generations of GM soybeans based on their cp4-epsps gene conserved se-quences. A widely adaptive detection method for the 1st and 2nd generation of cp4-epsps GM soybeans was established. Eva-luation on the method was conducted from the perspectives of accuracy, specificity, sensitivity and repeatability. ResultThe primer/probe was widely adaptive for detecting the 1st and 2nd generation of cp4-epsps GM soybeans. The detection method could detect cp4-epsps gene of 5 copies in reaction system, Ct was 38.88, and it could detect two generations at the same time. Accuracy and specificity evaluation indicated that only two generations of cp4-epsps GM soybeans could be detected. Forty-repeated test showed that the detection rate for 5×100 copies cp4-epsps gene fragment was 100%. Conclu-sionThe established detection method for the 1st and 2nd generation of cp4-epsps GM soybeans was of high specificity, sen-sitivity, repeatability, andaccuracy, which can be applicable to the monitoring of cp 4-epsps GM soybean and related products.

     

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