Figure 5.
The sRNA rnTrpL destabilizes the polycistronic trpDC mRNA. At an OD600nm of 0.5, one half of a liquid culture of strain 2011 ΔtrpL (pSRKTc-rnTrpL) was supplemented with IPTG for 10 min to induce lacZ‘-rnTrpL production, while the other half was incubated without IPTG. Then, rifampicin was added to both cultures and RNA was isolated at time points 0, 2, 4 and 6 min after rifampicin addition. Relative mRNA levels in induced (blue) and uninduced cultures (red) were determined by qRT-PCR analysis and half-lives (t1/2, in min) were calculated from three independent experiments, each with two technical replicates. The analysis was performed with primers specific for trpD (A), trpC (B) and trpE (C). The analysis with ppiD-, moaC- and moeA-specific primers is shown in Supplementary Figure S4.