Event-specific qualitative and quantitative detection of transgenic rice Kefeng-6 by characterization of the transgene flanking sequence

文献类型： 外文期刊

第一作者： Wang, Wei-Xia

作者： Wang, Wei-Xia;Lai, Feng-Xiang;Fu, Qiang;Zhu, Ting-Heng

作者机构：

关键词： Transgenic rice;Kefeng-6;Event-specific PCR;TAIL-PCR;Real-time PCR

期刊名称：EUROPEAN FOOD RESEARCH AND TECHNOLOGY （ 影响因子：2.998； 五年影响因子：3.005 ）

ISSN： 1438-2377

年卷期： 2011 年 232 卷 2 期

页码：

收录情况： SCI

摘要： Using TAIL-PCR (thermal asymmetric interlaced PCR), we analyzed the flanking sequences of transfer DNA (T-DNA) in transgenic rice Kefeng-6, which has insect-resistant genes, Cry1Ac and CpTi. Two junction sequences of T-DNA were identified in the Kefeng-6 genome: one integrated in chromosome 6 with an additional 22-bp insertion and another one in chromosome 9 with a 4-bp insertion between rice genomic DNA and T-DNA. Based on these inserts and border sequences, the event-specific qualitative and quantitative PCR system was established for this line. The relative limit of qualitative detection was assessed to be between 0.1 and 0.05%, and the absolute limit of detection in the quantitative PCR was approximately five initial copies. Two mixed rice samples with known Kefeng-6 contents were used to verify the quantitative method, from which the expected results were observed. This study provides a reliable method and information for detection, identification, and quantification of the presence of non-authorized GM rice Kefeng-6.

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