Københavns Universitet

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Master thesis project - Mode-of-action of novel phage endolysin at Københavns Universitet

Introduction: Phage-encoded endolysins are promising antimicrobials due to their efficient enzymatic activity towards the essential peptidoglycan of bacteria. We have identified an endolysins with a novel architecture, activity and regulation in phages of the Cp220viruses infecting Campylobacter jejuni.

Aim: Determine the mode-of-action and regulation of a novel endoIysin identified in the Cp220viruses infecting C. jejuni. Furthermore, you will identify remaining lytic genes such as the holin and the spanin and determine the regulation of all lytic proteins encoded by Cp220viruses.

Your work: You will investigate the mode-of-action and regulation of a novel endolysin in the Cp220virus phages using plasmid constructs containing the phage gene, and investigate its role by expression the phage protein in E. coli. You will make mutant constructs deleting various part of the endolysin to determine the role of the different domains of the protein and to determine the enzymatic active site. You will furthermore identify the holin in Cp220virus phages by co-expressing already identified holin candidates with the endolysin in E. coli. Furthermore, you will learn how to work with pathogenic bacteria and get GMOII lab experience as well as be working in an international research lab.

Methods: Mutant construction, protein expression, lytic assays, SDS page, bioinformatics etc.

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