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Identification of T-DNA Insertion Site and Flanking Sequence of a Genetically Modified Maize Event IE09S034 Using Next-Generation Sequencing Technology

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Abstract

Molecular characteristics including information of insertion site, flanking sequence, and copy numbers are the base for the safety assessment and subsequent monitoring of genetically modified organisms (GMOs), which has to be revealed thoroughly in a case-by-case manner. Although both polymerase chain reaction (PCR)-based and next-generation sequencing (NGS)-based approaches are proven to be effective in the molecular characterization of most of GM events, they often fail to work with GM maize events, mainly due to the genome complexity. In this study, by using NGS, we successfully identified the 3′ end T-DNA insertion site and flanking sequence of a GM maize event IE09S034, which were confirmed by PCR amplification and Sanger sequencing. Notably, insertions of unintended exogenous elements were revealed in this event although the single copy of target exogenous genes was also confirmed by digital PCR. The output of this study provides novel and important genetic evidence for the safety assessment and monitoring of GM maize event IE09S034.

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Acknowledgements

This work was supported by grants from the China National Transgenic Plant Special Fund (2016ZX08012-002, 2017ZX08013001-001) and the Programme of Introducing Talents of Discipline to Universities (111 Project, B14016).

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Authors and Affiliations

  1. Joint International Research Laboratory of Metabolic and Developmental Sciences, National Center for Molecular Characterization of Genetically Modified Organisms, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China

    Kiran Siddique, Jiaojun Wei, Rong Li, Dabing Zhang & Jianxin Shi

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  1. Kiran Siddique
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  2. Jiaojun Wei
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Contributions

JS conceived and designed the experiments. KS, JW, and RL performed the experiments. KS, RL, DZ, and JS analyzed and interpreted data. KS and JS wrote the manuscript.

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Correspondence to Jianxin Shi.

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Siddique, K., Wei, J., Li, R. et al. Identification of T-DNA Insertion Site and Flanking Sequence of a Genetically Modified Maize Event IE09S034 Using Next-Generation Sequencing Technology. Mol Biotechnol 61, 694–702 (2019). https://doi.org/10.1007/s12033-019-00196-0

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