title: Heterologous expression of drug-metabolizing enzymes in cellular and whole animal models.
abstract: In this report we describe the heterologous expression of glutathione S-transferase (GST) and cytochrome P450 reductase (Red) in E. coli and Salmonella typhimurium. The same expression vectors could be applied to both systems and high levels of catalytically active GST and Red were obtained. Interestingly the level of expression was invariably higher in S. typhimurium. The level of the alpha class GST being up to 20% of the total bacterial protein. A further advantage of the salmonella system is that strains were used which can be applied to mutagenicity tests. This system was validated by demonstrating increasing mutation frequency of halogenated hydrocarbons in strains expressing the GST and increased cytotoxicity of mitomycin C in cells expressing P450 reductase.
authors.affiliation: Imperial Cancer Research Fund, Ninewells Hospital, Dundee, UK.
authors.forename: A P
authors.initials: AP
authors.lastname: Simula
authors.forename: M B
authors.initials: MB
authors.lastname: Crichton
authors.forename: S M
authors.initials: SM
authors.lastname: Black
authors.forename: S
authors.initials: S
authors.lastname: Pemble
authors.forename: H F
authors.initials: HF
authors.lastname: Bligh
authors.forename: J D
authors.initials: JD
authors.lastname: Beggs
authors.forename: C R
authors.initials: CR
authors.lastname: Wolf
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D003001:Cloning, Molecular; D004926:Escherichia coli; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009152:Mutagenicity Tests; D009251:NADPH-Ferrihemoprotein Reductase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D011994:Recombinant Proteins; D012441:Saccharomyces cerevisiae; D012486:Salmonella typhimurium
chemical_list: D004364:Pharmaceutical Preparations; D011994:Recombinant Proteins; D009251:NADPH-Ferrihemoprotein Reductase; D005982:Glutathione Transferase