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. 2011 Jun 17;39(17):7586–7597. doi: 10.1093/nar/gkr477

Figure 4.

Figure 4.

Swapping of HCC region between ScoRsrA and BsuRsiW. (A) Amino acid sequences and secondary structure predictions were shown for B. subtilis RsiW (BsuRsiW) and ScoRsrA. The swapped regions (K33 to K47 in ScoRsrA and V26 to H40 in BsuRsiW) were indicated. (B) The S1 mapping results of swapped ScoRsrA and T-RsiW (truncated soluble form of BsuRsiW). In B. subtilis, the Tw32Δ2 (T-RsiW) and swapped T-RsiW (ST-RsiW) strains were grown to OD600 of 0.7 in LB and harvested at 0, 10, 20, 40, 80 min after diamide (1 mM) treatment. Transcripts from sigW promoters were analyzed by S1 mapping. Quantified values of the sigW transcript bands were normalized to ribosomal RNAs in each sample, and the relative expression values from three independent experiments were presented in comparison with the level in the absence of RsiW induction by IPTG. (C) In S. coelicolor, strains that contain wild-type rsrA (MK2) or swapped rsrA (S-RsrA) in the same genetic background (Supplementary Table S1) were grown to OD600 of ∼0.3 in YEME, and were harvested at 0, 10, 20, 40, 80, 120 min after treatment with 0.5 mM diamide. Transcripts from sigRp2 promoter were analyzed by S1 mapping. Quantified value of the SigR-dependent sigRp2 signal was normalized to the constitutive sigRp1 level in each sample. Relative expression values from more than three independent experiments were presented in comparison with the un-induced level.