Iron acquisition in Rhodococcus erythropolis: The isolation of mutant(s) that do not produce a siderophore

Rhodococcus, a soil bacterium, displays a diverse range of metabolic capabilities with a number of potential practical applications. To exploit the metabolic potential of Rhodococcus, their basic physiology, genetics, and especially the acquisition of essential nutrients like iron, must be understood.; R. erythropolis strain IGTS8 releases a small compound called a siderophore that scavenges ferric iron from the environment. To learn more about the genetic control of iron acquisition, mutant(s) defective in siderophore production were isolated. Mutants were generated by inserting a mutagenic plasmid, pJCS506, into the bacterial cell using electroporation. The plasmid, which cannot replicate in these bacterial cells, randomly inserts into the R. erythropolis genome producing mutations. The potential mutants were detected by screening on a chrome azurol S plate, which detects siderophore production. Colonies that failed to produce siderophore were tested by liquid assays. The strain N5-59 was confirmed as a non-siderophore producing mutant by liquid assays.