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.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Aspartate-Semialdehyde Dehydrogenase / genetics
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Bacterial Proteins / genetics
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Cloning, Molecular
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Escherichia coli / genetics
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Gene Expression Regulation, Bacterial / genetics*
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Genes, Reporter / genetics
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Genetic Markers / genetics
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Genetic Vectors / genetics*
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Gram-Negative Bacteria / enzymology
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Gram-Negative Bacteria / genetics*
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Green Fluorescent Proteins
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Lac Operon / genetics
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Luminescent Proteins / genetics
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Plasmids / genetics
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Promoter Regions, Genetic / genetics
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Pseudomonas aeruginosa / enzymology
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Pseudomonas aeruginosa / genetics*
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Sequence Homology, Amino Acid
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beta-Galactosidase / genetics
Substances
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Bacterial Proteins
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Genetic Markers
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Luminescent Proteins
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Green Fluorescent Proteins
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Aspartate-Semialdehyde Dehydrogenase
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beta-Galactosidase