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Investigating Transgene Integration and Organization in Cotton (Gossypium hirsutum L.) Genome

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Transgenic Cotton

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1902))

Abstract

In this chapter, we present detailed experimental procedures for investigating integration patterns of transgenes in cotton genome. We use conventional PCR and genomic Southern blot hybridization to characterize integration of T-DNA components and vector backbone fragments. For multiple-copy insertions into the same site (complex loci), transgene/transgene junctions (including canonical and truncated T-DNA and transgene involved vector backbone sequences) are characterized by PCR and sequencing. Inverse PCR and sequencing are used to characterize transgene/cotton genome junctions. Distribution of T-DNA insertion in cotton genome is evaluated by analysis of transgene flanking sequences. The pre-insertion sites can also be cloned and sequenced (based on the flanking sequences) for survey of genomic structure changes brought by transgene integration by comparing a pre-insertion site with corresponding transgene/plant junctions.

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Author information

Authors and Affiliations

  1. Cotton Research Center, Shandong Academy of Agricultural Sciences, Jinan, Shandong, P. R. China

    Jun Zhang

  2. School of Biological Sciences, Nanyang Technological University, Singapore, Singapore

    Yan Hong

Authors
  1. Jun Zhang
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  2. Yan Hong
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Corresponding author

Correspondence to Yan Hong .

Editor information

Editors and Affiliations

  1. Department of Biology, East Carolina University, Greenville, NC, USA

    Baohong Zhang

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Zhang, J., Hong, Y. (2019). Investigating Transgene Integration and Organization in Cotton (Gossypium hirsutum L.) Genome. In: Zhang, B. (eds) Transgenic Cotton. Methods in Molecular Biology, vol 1902. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8952-2_10

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  • DOI: https://doi.org/10.1007/978-1-4939-8952-2_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8951-5

  • Online ISBN: 978-1-4939-8952-2

  • eBook Packages: Springer Protocols

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