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Size Does Matter: Cre-mediated Somatic Deletion Efficiency Depends on the Distance Between the Target lox-Sites

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Abstract

Cre/lox recombination in vivo has become an important tool to induce chromosomal rearrangements like deletions. Using a combination of Ds transposition and Cre/lox recombination in two independent experiments on chromosomes 6 and 7 of tomato, two sets of somatic deletions up to a size of 200 kb were obtained. The efficiency of somatic deletion decreased with increasing deletion size. The largest germinally transmitted deletion had a size of only 55 kb. The results show that Cre-mediated deletion in somatic cells is less efficient when the lox sites are separated over larger distances. A further drop of the deletion efficiency after germinal transmission of the larger deletions can be explained by the probable loss of genes that are of vital importance to gametophyte function. Plasmid rescue of an 8.4 kb circularised deleted DNA showed that the Cre-mediated deletion takes place in tomato as expected. Since the circular Cre-deleted DNA could only be PCR amplified in plant cells where the deletion was not complete, the double-stranded DNA circle is assumed to be instable.

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Author notes

  1. Eric R. Coppoolse

    Present address: Laboratory of Bioinformatics, Wageningen University and Research Centre, Dreijenlaan 3, 6703, HA Wageningen, The Netherlands

Authors and Affiliations

  1. Department of Genetics, Faculty of Earth and Life Sciences, Institute for Molecular Cell Biology, Experimental, Vrije Universiteit, De Boelelaan 1087, 1081, HV Amsterdam, The Netherlands

    Eric R. Coppoolse, Marianne J. de Vroomen, Femke van Gennip & Bart J. M. Hersmus

  2. Plant Pathologie, Faculty of science, Swammerdam Institute for Life Sciences, University of Amsterdam, Kruislaan 318, 1098, SM Amsterdam, The Netherlands

    Marianne J. de Vroomen

  3. KeyGene N.V., AgroBusiness Park 90, P. O. box 216, 6700 AE, Wageningen, The Netherlands

    Mark J. J. van Haaren

Authors
  1. Eric R. Coppoolse
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  3. Femke van Gennip
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  4. Bart J. M. Hersmus
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  5. Mark J. J. van Haaren
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Coppoolse, E.R., de Vroomen, M.J., van Gennip, F. et al. Size Does Matter: Cre-mediated Somatic Deletion Efficiency Depends on the Distance Between the Target lox-Sites. Plant Mol Biol 58, 687–698 (2005). https://doi.org/10.1007/s11103-005-7705-7

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  • DOI: https://doi.org/10.1007/s11103-005-7705-7

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