Creating Genetic Engineering Tools for Investigating Bacillus anthracis.

Bacillus anthracis is a Gram positive, spore forming, non-motile, rod shaped, soil bacterium, and is endemic worldwide. Currently, the biology of B. anthracis is poorly understood. B. anthracis is one of many biological weapons used today. A -/- mutant strain of B. anthracis that lacks the pathogenic plasmids was created by serial culture at 42°C. Key DNA replication genes were identified by homology as targets. The dnaB gene, essential for B. subtilis initiation of DNA replication, was my focus. A vector system was created by polymerase chain reaction (PCR) with the pMUTIN4 integration vector and the promoter region of dnaB to study the genetics of B. anthracis. An electro-transformation system was formulated to knock-out the -/- B. anthracis dnaB gene. We have successfully incorporated the pMUTIN4 vector into the chromosomal DNA of B. anthracis. We also have formulated an electro-transformation system and vector system for use in B. anthracis