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. 2014 Mar 26;9(3):e93141. doi: 10.1371/journal.pone.0093141

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This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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(A) A schematic diagram of the transcriptional fusion of the rpoS promoter and 5’ untranslated region (UTR) fused directly to the lacZ _Bb open reading frame (ORF). The position of the rpoS transcriptional start site [58], [63] is indicated by a filled arrowhead. The Shine-Dalgarno sequence (RBS), and the translational start site of β-galactosidase (lacZ_Bb), indicated by the ATG, are also shown. Regions corresponding to the rpoS promoter and 5’ UTR (141bp) or the lacZ _Bb ORF are indicated with brackets. The "G" in the RBS marked with an asterisk is identified because the reporter construct harbored an A->G mutation relative to the published sequence at that position. Cell lysates from strains B31-S9 (wt), B31-S9/pBSV2G-rpoSp₁₄₁-lacZ_Bb (wt/rpoSp-lacZ_Bb), and B31-S9/pBSV2G- rpoSp₁₄₁-lacZ_Bb/pBSV28-flaBp-bbd18 (wt/rpoSp-lacZ_Bb/flaBp-bbd18) were grown under rpoS-inducing conditions and analyzed with OspC antisera (B) or stained with Coomassie blue (C) to demonstrate equivalent protein loads in each lane. The positions of molecular mass standards are shown on the left in kiloDaltons (kDa). (D) β-galactosidase activity in cell lysates from strains B31-S9 (wt), B31-S9/pBSV2G-rpoSp₁₄₁-lacZ_Bb (wt/rpoSp-lacZ_Bb), and B31-S9/pBSV2G- rpoSp₁₄₁-lacZ_Bb/pBSV28-flaBp-bbd18 (wt/rpoSp-lacZ_Bb/flaBp-bbd18) grown under rpoS-inducing conditions.