Evaluation of real-time PCR detection methods for detecting rice products contaminated by rice genetically modified with a CpTI-KDEL-T-nos transgenic construct
Evaluation of real-time PCR detection methods for detecting rice products contaminated by rice genetically modified with a CpTI-KDEL-T-nos transgenic construct
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DOI：
10.1016/j.foodchem.2013.05.005
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发表时间：
2013-12-01
期刊：
影响因子：
8.8
通讯作者：
Teshima, Reiko
中科院分区：
文献类型：
--
作者：
Nakamura, Kosuke;Akiyama, Hiroshi;Teshima, Reiko
转基因水稻（Oryza sativa）品系，如杀虫的克丰和克明稻，在许多国家被发现未经批准用于加工稻米产品。因此，转基因大米的定性检测方法是转基因食品监管的需要。在部分进口加工米制品中检测到一种表达豇豆胰蛋白酶抑制剂（CpTI）的转基因载体。所鉴定的用于表达CpTI的转基因构建体的3'末端序列包括内质网滞留信号编码序列（KDEL）和胭脂碱合酶终止子（T-nos）。该序列与科丰的一份报告中的序列相同。建立了一种新的构建体特异性实时聚合酶链反应（PCR）检测CpTI-KDEL和T-nos连接区序列的方法。采用构建体特异性实时荧光PCR方法对进口加工米产品进行转基因大米污染检测，并与5种事件特异性检测方法进行检测频率比较。构建体特异性检测方法检测转基因水稻的频率高于事件特异性检测方法。因此，我们建议构建体特异性检测方法是一个有益的工具，筛选转基因水稻品系，如科丰，在加工大米产品的转基因食品监管的污染。(C)2013爱思唯尔有限公司保留所有权利。
Genetically modified (GM) rice (Oryza sativa) lines, such as insecticidal Kefeng and Kemingdao, have been developed and found unauthorised in processed rice products in many countries. Therefore, qualitative detection methods for the GM rice are required for the GM food regulation. A transgenic construct for expressing cowpea (Vigna unguiculata) trypsin inhibitor (CpTI) was detected in some imported processed rice products contaminated with Kemingdao. The 3' terminal sequence of the identified transgenic construct for expression of CpTI included an endoplasmic reticulum retention signal coding sequence (KDEL) and nopaline synthase terminator (T-nos). The sequence was identical to that in a report on Kefeng. A novel construct-specific real-time polymerase chain reaction (PCR) detection method for detecting the junction region sequence between the CpTI-KDEL and T-nos was developed. The imported processed rice products were evaluated for the contamination of the GM rice using the developed construct-specific real-time PCR methods, and detection frequency was compared with five event-specific detection methods. The construct-specific detection methods detected the GM rice at higher frequency than the event-specific detection methods. Therefore, we propose that the construct-specific detection method is a beneficial tool for screening the contamination of GM rice lines, such as Kefeng, in processed rice products for the GM food regulation. (C) 2013 Elsevier Ltd. All rights reserved.