This thesis provides an overview of DNA replication proteins that potentially may serve as targets for antimicrobials in drug-resistant pathogens and includes the in silico identification of the... Show moreThis thesis provides an overview of DNA replication proteins that potentially may serve as targets for antimicrobials in drug-resistant pathogens and includes the in silico identification of the replication proteins of C. difficile. Additionally, the mode of action and the current developmental status of the compounds directed at the core replication machinery and accessory replication proteins are discussed (Chapter 2). In chapter 3 we determined the complete genome sequence of our reference laboratory strain, C. difficile 630Δerm, and compared this sequence to the sequence of C. difficile 630. This revealed an unexpected transposition of the mobile genetic element CTn5. The genome sequence now offers an appropriate reference for all molecular work on this strain. Apart from the identification and characterization of gyrase and DNA polymerase in previous studies, no information on DNA replication proteins or the mechanism of replication was available. To address this hiatus, we sought out to identify core replication proteins and to experimentally validate the functional role of these proteins. The findings of this experimental work, with a focus on helicase and primase, are presented in chapter 4 and chapter 5, respectively. Show less
