Supplemental Material to “ IpsA , a novel lacI - type regulator , is required for inositol - derived lipid formation in Corynebacteria and Mycobacteria ” , by Meike Baumgart

Growth experiments. For growth experiments, 5 ml of brain heart infusion broth (BHI, Difco Laboratories, Detroit, MI, USA) was inoculated with a colony from a fresh BHI agar plate and incubated for 6-8 h at 30°C and 220 rpm. Cells from this preculture were washed once in phosphate buffered saline (PBS, 137 mM NaCl, 2.7 mM KCl, 4.3 mM Na2HPO4, 1.4 mM KH2PO4, pH 7.3) and used to inoculate a second preculture, consisting of 20 ml CGXII minimal medium [1] supplemented with 3,4-dihydroxybenzoate (30 mg l -1 ) as an iron chelator and 2 % (w v -1 ) glucose as a carbon source, to an OD600 of about 1. For sequential cultivations, cells from the stationary phase were diluted in fresh medium to a starting OD600 of 1. After overnight incubation at 30°C and 120 rpm the main cultures were inoculated to an OD600 of about 1 and incubated at 30°C and 1200 rpm in a Biolector (m2p-labs, Baesweiler, Germany) in 48-well FlowerPlates containing 750 μl CGXII minimal medium and different carbon sources, as specified in the text. For induction of the expression of genes under the control of the Ptac-promoter, isopropyl β-D-1-thiogalactopyranoside (IPTG) was used at the concentrations specified. Where appropriate, the medium was supplemented with 25 μg l -1