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. 2014 Aug 26;9(8):e105644. doi: 10.1371/journal.pone.0105644

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© 2014 Santana 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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The 86-028NPrpsLΔfur(pT), 86-028NPrpsLΔfurΔhfq(pT), and 86-028NPrpsLΔfurΔhfq(pT-hfq) strains were grown to mid-logarithmic phase in DIS at 37°C with shaking at 50rpm. Rifampicin was then added to a final concentration of 250 µM to inhibit transcription. At 0, 2, 4, 6, 10, 20, and 30 minutes after rifampicin addition total RNA was collected. RNA was probed via northern blot with an RNA probe specific for the 5′ end of HrrF. Blots were stripped and probed again with an RNA probe specific for 5S rRNA to serve as a loading control (data not shown). The calculated half-life of HrrF was approximately seven minutes in the 86-028NPrpsLΔfur strain. There was no significant difference in the calculated HrrF half-life in any of the strains tested. This indicated that the stability of hrrF was not dependent on the RNA chaperone Hfq.