EspM regulates gene expression in response to the ESX-1 system in Mycobacterium marinum

Regulation of M. marinum gene expression in an espM deletion strain M Marinum M based strains (Wild type, epsM mutant and complemented overexpressor strain) were cultured in 5mL 7H9 and 0.1% Tween-80 for two days, then moved to 25mL 7H9 and 0.1% Tween-80 for two days. All strains were then subcultured in Sauton’s media 0.01% Tween-80 at an OD of 0.8  for 48 hours. The cultures were collected by centrifugation and stored at -80 for 24 hours. Cells were lysed using a bead beater. Total RNA was prepared from the pellets using the Qiagen RNeasy kit.