title: A Salmonella-based, propionate-inducible, expression system for Salmonella enterica.
abstract: The expression and regulatory properties of a propionate-regulated overexpression system (Salmonella enterica prpBCDE promoter (P(prpB)) and positive regulator (prpR)) were evaluated in wild-type S. enterica serovar Typhimurium TR6583 and prpB(-) or prpD(-) versions of this strain and compared with the arabinose-regulated T7 expression system. The wild-type strain showed low expression in the absence of propionate and high expression in the presence of propionate under all growth conditions. In 96-well plates and culture tubes, the wild-type strain exhibited a long delay before full induction; the time delay was significantly shorter in shake flasks. The prpD(-) strain exhibited low expression in the presence of glucose, highly regulatable expression over a wide range of propionate concentrations, and, in contrast to the wild-type strain, fast induction to full expression under all growth conditions. In contrast, the prpB(-) strain showed very high background expression in both culture tubes and shake flasks.
authors.affiliation: Department of Chemical Engineering, University of California, Berkeley, 94720, USA.
authors.forename: Sung Kuk
authors.initials: SK
authors.lastname: Lee
authors.forename: Jay D
authors.initials: JD
authors.lastname: Keasling
mesh_terms: D001089:Arabinose / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics; D017123:Bacteriophage T7 / Q000201:enzymology / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005798:Genes, Bacterial / Q000187:drug effects; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D010749:Phosphoprotein Phosphatases / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D011422:Propionates / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D019779:Salmonella enterica / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D011422:Propionates; D011994:Recombinant Proteins; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; D001089:Arabinose; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; C442127:PrpB protein, Salmonella enterica; D010749:Phosphoprotein Phosphatases; C029658:propionic acid