Tuberculosis continues to be a major cause of mortality worldwide, killing about two million people every year. The emergence of drug resistant strains of the bacterium has hindered efforts to successfully control the pandemic. The main causative agent, Mycobacterium tuberculosis, employs several strategies that help it survive in host macrophages. Recent studies have shown that the eukaryotic-like serine/threonine protein kinase G (PknG), secreted by M. tuberculosis, can inhibit phagosome-lysosome fusion and mediate intracellular survivability. Through transposon-insertion mutagenesis, a Mycobacterium smegmatis knockout of the pknG gene was generated. This mutant was found to be hypersusceptible to erythromycin, an antimycobacterial agent. This finding suggests that pknG is involved in processes that affect the activity of antimicrobial agents. The objective of the project is to characterize the roles that this gene plays in mycobacterial physiology, which may give us insight to the drug susceptibility and pathogenicity of mycobacteria, including M. tuberculosis .