FabT_+or-Tween_S.pyogenesM28_Transcriptomic analysis
FabT_+or-Tween_S.pyogenesM28_Transcriptomic analysis
GAS strains were cultured at 37 °C in THY or THY-Tween, and cells were harvested during exponential growth (OD600 between 0.4 and 0.5). Independent triplicate cultures were prepared for each condition.  For RNA preparation, 2 volumes of RNA protect* (Qiagen) was added to cultures prior centrifugation (10 min 12,000 g) and total RNA was extracted after lysing bacteria by a 30 min 15 mg.ml-1 lysozyme, 300 U.ml-1 mutanolysin treatment at 20 °C followed by two cycles of Fast-prep (power 6, 30 s) at 4 °C. RNA extraction (Macherey-Nagel RNA extraction kit; Germany) was done according to supplier instructions. RNA integrity was analyzed using an Agilent Bioanalyzer (Agilent Biotechnologies, Ca., USA). 23S and 16S rRNA were depleted from the samples using the MICROBExpress Bacterial mRNA enrichment kit (Invitrogen, France); depletion was controlled on Agilent Bioanalyzer (Agilent Biotechnologies).