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. 2011 Oct 11;6(10):e25705. doi: 10.1371/journal.pone.0025705

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This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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Uncut plasmids were separated by electrophoresis in agarose gels. Strain NCPPB 3335 (lane 1, WT) was mutagenized with Tn5-GDYN1 (8.8 kb) and insertions in plasmids pPsv48A (lane 7, A*), pPsv48B (lanes 2 and 4, B*) and pPsv48C (lane 6, C*) are evident by a retardation in mobility. Tagging or curing plasmid pPsv48B reveals the presence of plasmid pPsv48C (lanes 2, 4 and 5), which is of similar size but has a lower copy number. From the tagged derivatives, we obtained strains cured of plasmids pPsv48A (lanes 3 and 4, ΔA) and both pPsv48A and pPsv48B (lane 5, ΔAB). Lanes 6 and 7 correspond to strain B728a transformed with mutagenized plasmids pPsv48C and pPsv48A respectively. The molecular weights of the plasmids are indicated in kb to the left; lpc: Linearized plasmid and chromosomal DNA.