A promoter element upstream of nfsA likely drives expression of nfsA-H. A. Reverse transcriptase PCR analysis on the intergenic regions (indicated by lines in Figure 3) between nfsB and nsfC (B-C), nfsC and nfsD (C-D), nfsE and nfsF, (E-F), and nfsG and nfsH (G-H). PCR products were amplified from genomic DNA (gDNA) from wild type strain DK1622 (wt) or Δ(nfsA-H) strain PH1200 (nfs), and from cDNA generated in the absence (-RT) or presence (+RT) of reverse transcriptase, or from a mock cDNA reaction in which RNA was omitted (water). RNA from cells induced with glycerol for 0.5 hours was used to generate cDNA. B. Expression of mCherry reporter constructs during glycerol induced sporulation. Strains expressing mCherry fused after the promoter from pilA (PpilA::mCherry; strain PH1221), the 500 bp upstream of nfsA (PnfsA::mCherry; strain PH1220) or the vector backbone (vector; strain PH1222) were induced with glycerol, harvested at the indicated time points, and examined by fluorescence (left images) or DIC (right images) microscopy.
Müller et al. BMC Genomics 2010 11:264 doi:10.1186/1471-2164-11-264