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Frequent collinear long transfer of DNA inclusive of the whole binary vector during Agrobacterium-mediated transformation

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Abstract

When Agrobacterium was used to transform Nicotiana plumbaginifolia protoplasts and Arabidopsis thaliana roots and seedlings, a large number of plants were found in which not only the T-region defined by the border repeat sequences but the entire binary vector was integrated, as determined by both PCR and Southern analysis techniques. N. plumbaginifolia protoplast co-cultivation experiments yielded 3 out of 5 transformants with collinear sequence past the left border. In Arabidopsis root transformation experiments, 33% (6/18) of the transformants had T-DNA which exceeded the left border repeat. Vacuum infiltration of Arabidopsis seedlings produced even a greater percentage of transformants with sequences outside the left border repeat (62%, 39/63). The long transfer DNA cosegregated with the T-region encoded hygromycin resistance in the T2 progeny eliminating the possibility that long transfer DNA was of extrachromosomal or Agrobacterium origin. The high frequency of long transfer after vacuum infiltration of A. thaliana needs to be considered when analyzing T-DNA tagged mutants.

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Author notes

  1. Allan Wenck

    Present address: Forest Biotechnology Group, Dept. of Forestry, North Carolina State University, Box 8008, Raleigh, NC, 27695-8008, USA

  2. Ivan Kanevski

    Present address: Biomedical Consulting, 141 Birchview Drive, Piscataway, NJ, 08854, USA

Authors and Affiliations

  1. Department of Biology, University of South Carolina, Columbia, SC, 29208, USA

    Allan Wenck, Mihály Czakó, Ivan Kanevski & László Márton

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  1. Allan Wenck
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Wenck, A., Czakó, M., Kanevski, I. et al. Frequent collinear long transfer of DNA inclusive of the whole binary vector during Agrobacterium-mediated transformation. Plant Mol Biol 34, 913–922 (1997). https://doi.org/10.1023/A:1005849303333

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