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Event-specific quantitative detection of genetically modified wheat B72-8-11 based on the 3′ flanking sequence

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Abstract

Various PCR technologies have been developed for the execution of genetically modified organism (GMO) labeling policies of which an event-specific PCR detection method based on the flanking sequence of exogenous inserted DNA is the key trend in GMO detection due to its high specificity. In this study, 3′ flanking sequence between the host wheat DNA and the exogenous integrated gene construct of pHMW1Dx5 vector in transgenic wheat B72-8-11 was revealed by means of adaptor PCR, thus the fragment with the length of 210 bp was obtained, including a 144 bp unknown wheat genome DNA sequence. The event-specific quantitative PCR primers were designed based upon the revealed 3′ flanking sequence, and a SYBR Green I real-time PCR assay was subsequently applied. In the quantitative SYBR Green I real-time PCR assay, the LOD and LOQ were 10 and 20 haploid genome copies, respectively. In addition, three mixed wheat B72-8-11 samples with known contents were detected using this established quantitative PCR system, and the ideal results indicated that the developed event-specific quantitative PCR detection method could be used for identification and quantification of B72-8-11 wheat and its derivates.

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (Grant No. J1210069 and Grant No. 31100959) and Doctor Scientific Research Initial Foundation of Northeast Agricultural University (Grant No. 2012RCB19). This is gratefully acknowledged.

Conflict of interest

None.

Compliance with Ethics Requirements

This article does not contain any studies with human or animal subjects.

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Authors and Affiliations

  1. Key Laboratory of Agricultural Biological Functional Genes, Northeast Agricultural University, Harbin, Heilongjiang, China

    Minghui Zhang, Yanbo Yu, Xuejun Gao, Kejian Zhang, Yanming Zhu & Bo Qu

  2. Detecting Center of Agricultural Genetically Modified Organism, Research Center of Life Science and Biotechnique, Northeast Agricultural University, Harbin, Heilongjiang, China

    Minghui Zhang, Yanbo Yu, Xuejun Gao, Kejian Zhang & Bo Qu

  3. Heilongjiang Entry-Exit Inspection and Quarantine Bureau, Mudanjiang, Heilongjiang, China

    Fengxia Luan

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  1. Minghui Zhang
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  2. Yanbo Yu
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  3. Xuejun Gao
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  4. Kejian Zhang
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  5. Fengxia Luan
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  6. Yanming Zhu
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  7. Bo Qu
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Correspondence to Bo Qu.

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Zhang, M., Yu, Y., Gao, X. et al. Event-specific quantitative detection of genetically modified wheat B72-8-11 based on the 3′ flanking sequence. Eur Food Res Technol 240, 775–782 (2015). https://doi.org/10.1007/s00217-014-2383-9

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  • DOI: https://doi.org/10.1007/s00217-014-2383-9

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