【摘要】：【目的】建立一種廣譜性鑒定第一、二代轉(zhuǎn)cp4-epsp基因大豆的檢測方法,為準確鑒定第一、二代轉(zhuǎn)cp4-epsps基因大豆及其產(chǎn)品提供技術(shù)支持�！痉椒ā扛鶕�(jù)第一、二代轉(zhuǎn)基因大豆的cp4-epsp基因保守序列設(shè)計一對能同時鑒定兩代轉(zhuǎn)基因大豆外源基因cp4-epsps的引物和探針,建立一種廣譜性鑒定第一、二代轉(zhuǎn)cp4-epsps基因大豆的方法,并從準確性、特異性、靈敏性及重復(fù)性4個方面對該方法進行評估�！窘Y(jié)果】設(shè)計的引物/探針對鑒定第一、二代轉(zhuǎn)cp4-epsps基因大豆及其產(chǎn)品具有廣譜性;建立的檢測方法能檢測到反應(yīng)體系中低至5×100拷貝的cp4-epsps基因,Ct為38.88,且能同時檢測出兩代轉(zhuǎn)基因大豆。準確性和特異性評估結(jié)果顯示僅兩代轉(zhuǎn)cp4-epsp基因的大豆能被檢測到;40次重復(fù)試驗結(jié)果顯示,反應(yīng)體系中5×100拷貝的cp4-epsps基因片段的檢出率為100%�！窘Y(jié)論】建立的第一、二代轉(zhuǎn)cp4-epsps基因大豆鑒定方法具有特異強、靈敏度高、重復(fù)性好、準確性高等特點,可用于轉(zhuǎn)cp4-epsp基因大豆及其產(chǎn)品的監(jiān)測。
[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.
【作者單位】： 成都市石室中學文廟校區(qū);甘孜州農(nóng)產(chǎn)品質(zhì)量安全中心;四川省農(nóng)業(yè)科學院分析測試中心;
【基金】：四川省財政創(chuàng)新能力提升工程項目(2016GXTZ-010)
【分類號】：S565.1

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1 劉吉燾;馬曉杰;狄佳春;陳旭升;;棉花草甘膦抗性基因CP4-EPSPS的初步定位[J];江蘇農(nóng)業(yè)學報;2013年03期

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