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Transfection of heteroduplexes containing uracil · guanine or thymine · guanine mispairs into plant cells

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Abstract

We have compared the fate of U · G mispairs or analogous T · G mispairs in DNA heteroduplexes transfected into tobacco protoplasts. The heteroduplex DNA consisted of tomato golden mosaic virus DNA sequences in theEscherichia coli vectors pUC118 or pUC119. After transfection, the mismatched U residues were lost with an efficiency of greater than 95%, probably as a result of the uracil-DNA glycosylase pathway for excision of U residues in any sequence context. In contrast to the preferential removal of the mispaired U residues, biased removal of T residues from analogous heteroduplexes was not seen in the transfected plant cells. Also, we investigated the effect of extensively methylating one strand of the heteroduplex DNA used for transfection. Surprisingly, such methylation resulted in highly biased loss of the mismatched base from the 5-methylcytosine-rich strand of T · G-containing heteroduplexes.

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Author notes

  1. Clare L. Brough

    Present address: Department of Biological Sciences, University of Durham, DH1 5 EY, Durham, UK

Authors and Affiliations

  1. Department of Biochemistry, Tulane Medical School, 70112, New Orleans, LA, USA

    Nilufar M. Inamdar, Xian-Yang Zhang & Melanie Ehrlich

  2. Biotechnology Center, The Ohio State University, 43210, Columbus, OH, USA

    Clare L. Brough, William E. Gardiner & David M. Bisaro

  3. Department of Molecular Genetics, The Ohio State University, 43210, Columbus, OH, USA

    Clare L. Brough, William E. Gardiner & David M. Bisaro

Authors
  1. Nilufar M. Inamdar
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  2. Xian-Yang Zhang
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  3. Clare L. Brough
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  4. William E. Gardiner
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  5. David M. Bisaro
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  6. Melanie Ehrlich
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Deceased. We dedicate this paper to the memory of this young scientist.

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Inamdar, N.M., Zhang, XY., Brough, C.L. et al. Transfection of heteroduplexes containing uracil · guanine or thymine · guanine mispairs into plant cells. Plant Mol Biol 20, 123–131 (1992). https://doi.org/10.1007/BF00029155

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  • DOI: https://doi.org/10.1007/BF00029155

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