Genome wide analysis of the GlnR regulon in Mycobacterium smegmatis
Genome wide analysis of the GlnR regulon in Mycobacterium smegmatis
M. smegmatis strains were grown in triplicate in nitrogen limiting conditions as stated until external nitrogen had completely depleted. Cells were immediately added to final concentration 2.5 M GTC and harvested by centrifugation. Pellets were resuspended in 1 ml TRIzol (Life Technologies) and stored at -80 C. Thawed cell/Trizol suspensions were transferred to tubes containing Zirconium beads (MP Biomedicals) and the cells lysed in a ribolyser Fastprep (Hybaid). RNA samples were chloroform extracted and RNA purified using the RNeasy kit (Qiagen) according to manufacturer's instructions.
Experimental Factor: IMMUNOPRECIPITATE
INPUT
Experimental Factor: compound
ammonium sulphate nitrogen source
Experimental Factor: dose
1 millimolar