| 第一、二代转cp4-epsps基因大豆鉴定方法的建立 |
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| 引用本文： | 宋秋池,刘熹薇,牟益位,敬媛,宋君.第一、二代转cp4-epsps基因大豆鉴定方法的建立[J].南方农业学报,2017,48(7). |
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| 作者姓名： | 宋秋池 刘熹薇 牟益位 敬媛 宋君 |
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| 作者单位： | 1. 成都市石室中学 文庙校区,成都,610091;2. 甘孜州农产品质量安全中心,四川 康定,626000;3. 四川省农业科学院 分析测试中心,成都,610066 |
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| 基金项目： | 四川省财政创新能力提升工程项目 |
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| 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。
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| 关 键 词： | 转基因大豆 cp4-epsps基因 广谱性 特异性 灵敏度 |
Establishment of detection method for the 1st and 2nd generation of cp4-epsps genetically modified soybeans |
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| SONG Qiu-chi,LIU Xi-wei,MOU Yi-wei,JING Yuan,SONG Jun.Establishment of detection method for the 1st and 2nd generation of cp4-epsps genetically modified soybeans[J].Journal of Southern Agriculture,2017,48(7). |
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| Authors: | SONG Qiu-chi LIU Xi-wei MOU Yi-wei JING Yuan SONG Jun |
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| Abstract: | Objective]A 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. Method]A 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. Result]The 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-sion]The 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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| Keywords: | genetically modified soybean cp4-epsps gene wide adaptability specificity sensitivity |
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