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Multiplex event-specific qualitative polymerase chain reaction for detecting three transgenic rice lines and application of a standard plasmid as a quantitative reference molecule

文献类型： 外文期刊

作者： Wang, Xiaofu 1 ; Chen, Xiaoyun 2 ; Xu, Junfeng 2 ; Wang, Pengfei 3 ; Shen, Wenbiao 1 ;

作者机构： 1.Nanjing Agr Univ, Coll Life Sci, Nanjing 210095, Jiangsu, Peoples R China

2.Zhejiang Acad Agr Sci, Inst Qual & Standard Agr Prod, Hangzhou 310021, Zhejiang, Peoples R China

3.Shenyang Normal Univ, Coll Chem & Life Sci, Shenyang 110034, Peoples R China

关键词： Genetically modified rice;Multiplex PCR;Plasmid;TT51-1;KMD1;KF6

期刊名称：ANALYTICAL BIOCHEMISTRY （ 影响因子：3.365； 五年影响因子：3.049 ）

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收录情况： SCI

摘要： The three most well-known genetically modified (GM) rice lines in China are TT51-1, KMD1, and KF6. The purposes of this study were to establish a multiplex event-specific qualitative polymerase chain reaction (meqPCR) system for simultaneous detection of the three transgenic rice events and to construct a plasmid as the reference molecule for quantitative analysis. Event-specific primers for each event were selected or designed by focusing on the transgene borders between the inserted DNA and the flanking rice DNA. The developed meqPCR was anticipated to detect distinct amplicons as 454, 398, 301, and 250 bp from KF6, KMD1, TT51-1, and the rice endogenous reference gene, respectively. The robustness of the meqPCR was tested with different levels of the three transgenic rice genomic DNAs, and the sensitivity threshold of the meqPCR was at least 50 ng of 0.1% rice DNA for each event when the three transgenic rice events present and with other GM materials together. The constructed plasmid was evaluated using mixed samples with known GM contents in real-time quantitative PCR. The results indicated that the constructed plasmid was acceptable and suitable for GM rice quantitative analysis. (C) 2014 Elsevier Inc. All rights reserved.

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[1]Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification. Chen, Xiaoyun,Wang, Xiaofu,Zhou, Yu,Huang, Sainan,Miao, Qingmei,Zhu, Qing,Xu, Junfeng,Jin, Nuo,Huang, Sainan. 2012

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