title: Detection of induced synthesis of colicin E9 using ColE9p::gfpmut2 based reporter system.
abstract: The majority of colicin operons are regulated by an SOS response inducible promoter (SOS promoter), located at upstream of the colicin operons. Therefore, colicin synthesis is induced by DNA damaging agents like mitomycin C (MMC) because the resulting DNA damage switches on the SOS response in bacteria. In this study, we have described the strategy for fusion of the SOS promoter of the colicin E9 operon (ColE9p) with a promoterless green fluorescent reporter gene (gfpmut2). We observed that the ColE9p-gfpmut2 is inducible by MMC which confirmed that the ColE9p-gfpmut2 is sensitive to SOS response inducing agents. The data implies that the ColE9p-gfpmut2 based reporter system is suitable for monitoring the ColE9 synthesis induced by SOS response inducing agents including antibiotics. Using green fluorescent protein expression from the ColE9p-gfpmut2 as an indicator of ColE9 synthesis; we have investigated, first time, the inducing effects of cephalexin antibiotic on ColE9 synthesis. Our data demonstrated that the cephalexin has potential to induce ColE9 synthesis from E. coli JM83 host cells albeit the level of this induction is very low hence its detection required a highly sensitive method.
authors.affiliation: School of Life Sciences, Centre for Biomolecular Sciences, University of Nottingham, University Park, Nottingham, NG7 2RD, UK, shaista.bano@usindh.edu.pk.
authors.forename: Shaista
authors.initials: S
authors.lastname: Bano
authors.forename: Mireille
authors.initials: M
authors.lastname: Vankemmelbeke
authors.forename: Christopher N
authors.initials: CN
authors.lastname: Penfold
authors.forename: Richard
authors.initials: R
authors.lastname: James
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002506:Cephalexin / Q000494:pharmacology; D003087:Colicins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D009876:Operon / Q000235:genetics; D013014:SOS Response, Genetics / Q000235:genetics / Q000502:physiology
chemical_list: D000900:Anti-Bacterial Agents; D003087:Colicins; D029968:Escherichia coli Proteins; D002506:Cephalexin