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 β-semialdehyde dehydrogenase (asd) gene as a selectable marker and β-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.