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Biology 2014, 3(1), 39-55; doi:10.3390/biology3010039

Analysis of T-DNA/Host-Plant DNA Junction Sequences in Single-Copy Transgenic Barley Lines

Department of Crop Genetics, John Innes Centre, Norwich Research Park, Norwich, NR4 7UH, UK
Author to whom correspondence should be addressed.
Received: 4 November 2013 / Revised: 3 January 2014 / Accepted: 3 January 2014 / Published: 21 January 2014
(This article belongs to the Special Issue Insights from Plant Genomes)
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Sequencing across the junction between an integrated transfer DNA (T-DNA) and a host plant genome provides two important pieces of information. The junctions themselves provide information regarding the proportion of T-DNA which has integrated into the host plant genome, whilst the transgene flanking sequences can be used to study the local genetic environment of the integrated transgene. In addition, this information is important in the safety assessment of GM crops and essential for GM traceability. In this study, a detailed analysis was carried out on the right-border T-DNA junction sequences of single-copy independent transgenic barley lines. T-DNA truncations at the right-border were found to be relatively common and affected 33.3% of the lines. In addition, 14.3% of lines had rearranged construct sequence after the right border break-point. An in depth analysis of the host-plant flanking sequences revealed that a significant proportion of the T-DNAs integrated into or close to known repetitive elements. However, this integration into repetitive DNA did not have a negative effect on transgene expression. View Full-Text
Keywords: transgene; flanking sequence; junction sequence; T-DNA integration; transgenic barley transgene; flanking sequence; junction sequence; T-DNA integration; transgenic barley

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This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Bartlett, J.G.; Smedley, M.A.; Harwood, W.A. Analysis of T-DNA/Host-Plant DNA Junction Sequences in Single-Copy Transgenic Barley Lines. Biology 2014, 3, 39-55.

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