title: ASD-GFP vectors for in vivo expression technology in Pseudomonas aeruginosa and other gram-negative bacteria.
abstract: We describe the construction of promoter probe vectors designed for identification of bacterial genes induced in vitro and/or in vivo and for measurement of gene expression levels for in vivo expression technology. These plasmids use the Pseudomonas aeruginosa aspartate beta-semialdehyde dehydrogenase (asd) gene as a selectable marker and beta-galactosidase (pIVPRO, 10.88 kb) or mutant green fluorescent protein with enhanced fluorescence properties (mut3GFP, pIVET-GFP, 5.48 kb) as reporter gene systems. The proposed strategies can be adapted for use in most Gram-negative bacteria.
authors.affiliation: Université Laval, Ste.-Foy, QC, Canada.
authors.forename: M
authors.initials: M
authors.lastname: Handfield
authors.forename: H P
authors.initials: HP
authors.lastname: Schweizer
authors.forename: M J
authors.initials: MJ
authors.lastname: Mahan
authors.forename: F
authors.initials: F
authors.lastname: Sanschagrin
authors.forename: T
authors.initials: T
authors.lastname: Hoang
authors.forename: R C
authors.initials: RC
authors.lastname: Levesque
mesh_terms: D001223:Aspartate-Semialdehyde Dehydrogenase / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics; D005819:Genetic Markers / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006090:Gram-Negative Bacteria / Q000201:enzymology / Q000235:genetics*; D049452:Green Fluorescent Proteins; D007763:Lac Operon / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000201:enzymology / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D005819:Genetic Markers; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D001223:Aspartate-Semialdehyde Dehydrogenase; D001616:beta-Galactosidase