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. 2008 May 7;3(5):e2087. doi: 10.1371/journal.pone.0002087

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Farhana et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) msmeg_6554, a homologue of M.tb irtA in M.smeg was disrupted by introducing hygromycin cassette as described in materials and methods. A 3.3 kb insert at the msmeg_6554 locus was incorporated to create the knockout. (B) PCR amplifications indicating the disruption of msmeg_6554. Lane 2, negative control; lane 3, amplification from M.smeg mc²155 genomic DNA; lane 4, amplification from PCK48hyg vector; lane 5, KO48-SCO, single crossover; lane 6, KO48-DCO, double crossover. (C) RT-PCR of msmeg_6553 on RNA isolated from mc²155▵6554 grown under iron replete (lane 1) and iron depleted (lane 2) conditions.