Identifying growth deficient \u3ci\u3eBacillus anthracis\u3c/i\u3e mutants via transponson mutagenesis

Bacillus anthracis is a spore-forming bacterium and etiological agent of the disease anthrax. How exactly B. anthracis spores germinate and grow into their vegetative form is a poorly understood concept, and the bacterium\u27s metabolism and nutrient requirements within a host environment are largely unknown. Our aim was to start an investigation into the metabolic requirements of B. anthracis that help it to flourish so effectively in the host. The first step in this project was the creation of a B. anthracis mutant library. Creation of the library was facilitated by the use of transposon mutagenesis, where mobile transposons insert into the host genomic DNA causing a disruption of gene function. A library of nearly 400 mutants was generated and screened for growth impairment in minimal media using optical density measurements. Arbitrarily selected mutants with a defective growth phenotype were analyzed further by using a Southern Blot to confirm the presence of the transposon, indicating a disruption in a gene or genes necessary for effective growth. This library of mutants can be studied further in order to identity the metabolic pathways affected, thus revealing novel targets for therapy