The 168 genome contains an array of alternative σ factors, many of which play important roles in reprogramming expression during stress and sporulation. The role of the different σ factors during outgrowth, when the germinated endospore is converted back to a vegetative cell, is less well characterized. The activity of the alternative σ factors σ, σ and σ during endospore outgrowth was analysed by Northern blotting and reporter assays. While σ and σ were transcriptionally active during outgrowth, σ-dependent transcription was not observed until after the first cell division, when growth slowed. Using an IPTG-controllable copy of , an optimal level of expression was required to maintain growth rate at the end of outgrowth. The genes encoding the putative extracytoplasmic function (ECF) σ factors σ, σ, σ, σ and YlaC were insertionally inactivated using pMUTIN4. These strains, together with and mutants, were tested to determine their role and measure their expression during endospore outgrowth. Transcripts or β-galactosidase activity were observed for each of the ECF σ factors early after germination. With the exception of MJH003 (), which showed an exacerbated salt stress defect, inactivation of the ECF σ factor genes did not affect outgrowth in the conditions tested.


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