Comparison of Transcription Profiles of M. smegmatis overexpressing wild type and phopshorylation defective PknK

We have shown that elevated levels of wild-type M. tuberculosis PknK (PknK Mtb), but not phosphorylation-defective PknKMtb, in Mycobacterium smegmatis cause significant retardation of growth rate and altered colony morphology (Malhotra et al., 2012). LIX79 (WT PknK) and LIX80 (PknK K55M mutant) M. smegmatis strains will be grown in LB and induced with 0.2% acetamide for the induction of wild type or phosphorylation defective pknK gene. RNA and Protein will be isolated from cultures pelleted at times—24 h (24h) and 96 h (96 h) post induction. M. smegmatis strain LIX70 containing the empty vector will be induced at the same time-points and used as a control in hybridization. Two independent replicate experiments will be done. Two color Experiment,Organism: Mycobacterium smegmatis, Agilent Genotypic Technology designed Custom Mycobacterium smegmatis Whole Genome 8x15k GE Microarray (AMADID-43029)