Team:Oxford/Experiments

Team Oxford

One-paragraph summary of our technical plan The project aims to investigate how bacterial biofilm-disrupting proteins can be exported from E. coli. The proteins Dispersin B, Microcin S, DNase, and Artilysin will be expressed from commercial pBAD expression vectors with N-terminal fusion tags to target them for export via the DsbA (Sec), YebF (Porin), and flagellar export pathways in E. coli. Additionally, the holin gene will be expressed under the control of bacterial quorum sensing-responsive promoters (in conjunction with the artilysin) to cause host cell lysis and release of these proteins from the cytoplasm on detection of a target cell density. Protein secretion will be monitors by the presence of a His tag on the secreted proteins detected by Western blotting of the growth medium.