第一、二代转cp4-epsps基因大豆鉴定方法的建立
发布时间：2019-04-12 07:27
【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]:[objective] to establish a broad-spectrum identification method for the first and second generation of cp4-epsp transgenic soybean and provide technical support for the accurate identification of the first and second generation of soybean and its products. [methods] according to the first, the second generation of transgenic soybean and its products. A pair of primers and probes that can simultaneously identify the exogenous gene cp4-epsps of the second generation of transgenic soybean were designed to establish a broad spectrum identification method for the first and second generation of cp4-epsps transgenic soybean, and the cp4-epsp gene conserved sequence of the second generation transgenic soybean was designed to identify the foreign gene cp4-epsps of the two generation transgenic soybean at the same time. The method was evaluated from the aspects of accuracy, specificity, sensitivity and repeatability. [results] the primers / probes designed for the identification of the first and second generations of cp4-epsps-transgenic soybean and its products had broad spectrum. The established detection method can detect the cp4-epsps gene with a Ct of 38.88 and can detect two generations of transgenic soybean in the reaction system as low as 5 脳 100 copies. The results of accuracy and specificity evaluation showed that only two generations of soybean with cp4-epsp gene could be detected. The results of 40 repeated tests showed that the detection rate of 5 脳 100 copies of cp4-epsps gene fragments in the reaction system was 100%. [conclusion] the first and second generation cp4-epsps transgenic soybean identification methods have strong specificity, high sensitivity and good repeatability. With high accuracy, it can be used for the monitoring of cp4-epsp-transgenic soybean and its products.
【作者单位】： 成都市石室中学文庙校区;甘孜州农产品质量安全中心;四川省农业科学院分析测试中心;
【基金】：四川省财政创新能力提升工程项目(2016GXTZ-010)
【分类号】：S565.1
本文编号：2456830
[Abstract]:[objective] to establish a broad-spectrum identification method for the first and second generation of cp4-epsp transgenic soybean and provide technical support for the accurate identification of the first and second generation of soybean and its products. [methods] according to the first, the second generation of transgenic soybean and its products. A pair of primers and probes that can simultaneously identify the exogenous gene cp4-epsps of the second generation of transgenic soybean were designed to establish a broad spectrum identification method for the first and second generation of cp4-epsps transgenic soybean, and the cp4-epsp gene conserved sequence of the second generation transgenic soybean was designed to identify the foreign gene cp4-epsps of the two generation transgenic soybean at the same time. The method was evaluated from the aspects of accuracy, specificity, sensitivity and repeatability. [results] the primers / probes designed for the identification of the first and second generations of cp4-epsps-transgenic soybean and its products had broad spectrum. The established detection method can detect the cp4-epsps gene with a Ct of 38.88 and can detect two generations of transgenic soybean in the reaction system as low as 5 脳 100 copies. The results of accuracy and specificity evaluation showed that only two generations of soybean with cp4-epsp gene could be detected. The results of 40 repeated tests showed that the detection rate of 5 脳 100 copies of cp4-epsps gene fragments in the reaction system was 100%. [conclusion] the first and second generation cp4-epsps transgenic soybean identification methods have strong specificity, high sensitivity and good repeatability. With high accuracy, it can be used for the monitoring of cp4-epsp-transgenic soybean and its products.
【作者单位】： 成都市石室中学文庙校区;甘孜州农产品质量安全中心;四川省农业科学院分析测试中心;
【基金】：四川省财政创新能力提升工程项目(2016GXTZ-010)
【分类号】：S565.1
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1 刘吉焘;马晓杰;狄佳春;陈旭升;;棉花草甘膦抗性基因CP4-EPSPS的初步定位[J];江苏农业学报;2013年03期
，本文编号：2456830
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