Project description:The goal of this study is to obtain a genomic view of the Fur regulatory network under both iron replete and iron deficient conditions in Bacillus subtilis using ChIP-seq. Besides the known Fur target sites, 70 putative DNA binding sites were identified, and the vast majority had higher occupancy under iron sufficient conditions. In addition,we discovered a role for catechol degradation in bacillibactin metabolism, and provided evidence that catechol 2,3-dioxygenase can detoxify endogenously produced catechol substrates in addition to its more widely studied role in biodegradation of environmental aromatic compounds and pollutants. Overall design: To modulate intracellular iron levels, we employed a high-affinity Fe2+ exporter FrvA from L. monocytogenes to impose iron starvation. Bacillus wild-type cells (with C-terminal FLAG-tagged Fur at its native locus and an IPTG-inducible ectopic copy of frvA integrated at amyE locus) were harvested at 0 and 30 min after IPTG induction to study Fur-dependent regulation under iron sufficient and deficient conditions, respectively.
Project description:Comparison of the Bacillus cereus with overexpressed Bacillus subtilis ComK (Bacillus cereus pNWcomKBsu) vs Bacillus cereus carrying empty plasmid (Bacillus cereus pNW33N) One condition design comparision of (IPTG induced overexpression construct vs IPTG induced empty plasmid) including a dye swap, 3 biological replicate