ASD-GFP vectors for in vivo expression technology in Pseudomonas aeruginosa and other gram-negative bacteria.
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.
Handfield
M
M
Université Laval, Ste.-Foy, QC, Canada.
Schweizer
H P
HP
Mahan
M J
MJ
Sanschagrin
F
F
Hoang
T
T
Levesque
R C
RC
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
D001426:Bacterial Proteins; D005819:Genetic Markers; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D001223:Aspartate-Semialdehyde Dehydrogenase; D001616:beta-Galactosidase