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. 2011 Dec 16;40(8):3623–3640. doi: 10.1093/nar/gkr1156

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© The Author(s) 2011. Published by Oxford University Press.

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

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Figure 4. — (A) Pulse expression of SdsR sRNA from an inducible P_BAD promoter results in rapid decrease of ompD mRNA levels. Salmonella ΔsdsR and ΔsdsR ΔompD (JVS-8434) cells carrying either the control vector pBAD (pKP8-35) or the expression plasmid pBAD-SdsR (pKP19-8) were grown to an OD₆₀₀ of 1.5 and total RNA samples were collected prior to and at indicated timepoints after l-arabinose addition (0.2% final concentration). Both ompD mRNA and SdsR sRNA levels were detected by northern blotting. (B) SdsR requires RNase E for ompD mRNA decay. Salmonella rne-ctrl. deleted for rybB, micC, invR and sdsR sRNA genes (JVS-9549) and its isogenic rne-ts strain (JVS-9550) were transformed with pBAD-SdsR and grown at 30°C to an OD₆₀₀ of 1.5 when cultures were split. Growth was continued for 30 min at 30°C or, to inactivate RNase E in rne-ts, at 44°C prior to arabinose-induced expression of SdsR for 10 min. Levels of ompD mRNA, SdsR RNA and 5S RNA (loading control) were determined by northern blot analysis of total RNA.