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. 2013 Aug 31;41(22):10403–10413. doi: 10.1093/nar/gkt757

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© The Author(s) 2013. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — Inability of circular plasmids to replicate in the ΔparE mutants, M145ΔparE and 3456ΔparE. (A) Circular plasmids pLUS356 and pLUS891 (left) used for transformation and the linear versions, pLUS356L and pLUS891L, generated in the transformants (right). bla, beta-lactamase gene; tsr, thiostrepton resistance gene; ARS, autonomously replicating sequence of pSLA2; filled arrows, telomeres of the S. lividans chromosome (on pLUS356) or SCP1 plasmid (on pLUS891); filled circles, TPs. As, AseI site; Ba, BamHI site. The sizes (kb) of the BamHI fragments of the linear DNA are indicated. (B) M145ΔparE and 3456ΔparE were transformed by pLUS356 or pLUS891. Genomic DNA was isolated from the Thio^r transformants, digested with BamHI, and subjected to Southern blotting using the transforming DNA as the respective probes. Representative transformants are shown. The sizes (kb) of the hybridizing fragments are indicated.