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Event-specific detection of genetically modified wheat B73-6-1 based on the 3′-flanking sequence

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Abstract

In this study, 3′-flanking sequence between the host plant DNA and the integrated gene construct of pHMW1Dx5 vector in transgenic wheat B73-6-1 was revealed by means of adaptor PCR; thus, the fragment with the length of 3.1 kb was obtained, including a 190-bp wheat genomic DNA, which demonstrates that this HMW-GS gene was located on the wheat chromosome 3B. And the event-specific PCR primers were designed based upon the revealed 3′-flanking sequence; the conventional qualitative PCR and quantitative SYBR real-time PCR detection methods employing these primers were successfully developed. In conventional qualitative PCR assay, the limit of detection was 0.1 % for B73-6-1 wheat genomic DNA for one reaction. In the quantitative SYBR real-time PCR assay, the limit of detection and limit of quantification were 10 and 100 haploid genome copies, respectively. In addition, three mixed blind wheat samples with known B73-6-1 contents were detected using the established real-time PCR systems, and the ideal results indicated that the established event-specific real-time PCR detection systems were reliable, sensitive and accurate.

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Acknowledgments

This study was supported by the foundation of Educational Committee of Heilongjiang Province of China (No.12521018). This is gratefully acknowledged.

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Authors and Affiliations

  1. Detecting Center of Agricultural Genetically Modified Organism, Research Center of Life Science and Biotechnique, Northeast Agricultural University, 59 Mucai Road, Harbin, 150030, People’s Republic of China

    Minghui Zhang, Nan Huo, Ying Liu, Youwen Qiu, Jinxia Ao, Qingzhang Li & Xuejun Gao

  2. Heilongjiang Entry-Exit Inspection and Quarantine Bureau, Harbin, 150001, People’s Republic of China

    Fengxia Luan

Authors
  1. Minghui Zhang
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  2. Nan Huo
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  3. Ying Liu
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  4. Youwen Qiu
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  5. Jinxia Ao
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  7. Qingzhang Li
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  8. Xuejun Gao
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Correspondence to Xuejun Gao.

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Zhang, M., Huo, N., Liu, Y. et al. Event-specific detection of genetically modified wheat B73-6-1 based on the 3′-flanking sequence. Eur Food Res Technol 235, 1149–1159 (2012). https://doi.org/10.1007/s00217-012-1848-y

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  • DOI: https://doi.org/10.1007/s00217-012-1848-y

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