Effects of yhcN and ylaJ deletions during spore germination in Bacillus anthracis

Certain bacteria produce spores as a defense mechanism. Bacterial spores have thick walls, are very resistant, and remain inactive until they germinate. To complete germination, the spore needs to undertake a process of degrading the spore cortex peptidoglycan by the activity of germination-specific lytic enzymes (GSLEs). GSLEs can be subdivided into two categories: Spore Cortex Lytic Enzymes (SCLEs) and Cortex Fragment Lytic Enzymes (CFLEs). The Bacillus anthracis GSLEs are SleB, CwlJ1, CwIJ2 and SleL. Enzymes that play an important role in the degradation of the cortex so that germination can occur are SleB and CwIJ1. Bacillus anthracis has three genes, sleB, ypeB and ylaJ, in an operon that act as a unit of transcription. YlaJ is a sporulation-specific protein encoded in the operon. A similar gene, yhcN is found in the chromosome of B. anthracis and plays a small role in spore germination. In this study ΔyhcN ΔylaJ and ΔyhcN strains will be created in B. anthracis using the markerless gene replacement strategy. Spores of these deletions strains will then be analyzed for the germination rates and efficiency and for the activity of sleB in degrading the cortex. The objective of this study is to determine the specific function of these two genes during germination and also to determine the effect on spore when both genes are deleted. The deletion of these two genes of the spore represents the beginning and the hope of what later may be the cure of the disease that causes Bacillus anthracis, the anthrax.