Identifying the genome-wide binding sites of DnaA in Mycobacterium tuberculosis and Mycobacterium smegmatis

The purpose of this study was to identify the genome-wide binding sites of DnaA in Mycobacterium tuberculosis and Mycobacterium smegmatis. Two experiments were performed and are included in this project. First, in vitro DNA affinity sequencing (IDAP-seq) in which purified M. tuberculosis DnaA protein was used to pull down sheared M tuberculosis genomic DNA. In addition, two different point mutants of dnaA (I282T and R400H) were also used to perform IDAP-seq. The second experiment utilized a strain of Mycobacterium smegmatis in which dnaA has been deleted and a N-terminal myc-epitope tagged copy of dnaA was inserted at another site in the genome. ChIP-seq experiment was then performed on this strain and a strain bearing an untagged copy as a control.