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. 2015 Jan 23;14:11. doi: 10.1186/s12934-015-0194-8

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© Wang et al.; licensee BioMed Central. 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Schematics illustrating the use of the pK18 mobsacB -Cm or pK18 mobsacB -Ery suicide plasmids to generate a defined, marker-free deletion in Pseudoalteromonas strains. (A) Construction of suicide plasmid constructs containing the homologous fragments flanking the target DNA region. (B) First recombination event by integration into the chromosome via homologous recombination and position of the primers for plasmid insertion verification. (C) The second recombination event by sucrose selection, and position of primers to separate the deletion mutants from the wild-type.