M. Handfield et al., ASD-GFP VECTORS FOR IN-VIVO EXPRESSION TECHNOLOGY IN PSEUDOMONAS-AERUGINOSA AND OTHER GRAM-NEGATIVE BACTERIA, BioTechniques, 24(2), 1998, pp. 261-264
We describe the construction of promoter probe vectors designed for id
entification of bacterial genes induced in vitro and/or in vivo and fo
r measurement of gene expression levels for in vivo expression technol
ogy These plasmids use the Pseudomonas aeruginosa aspartate beta-semia
ldehyde dehydrogenase (asd) gene as a selectable marker and beta-galac
tosidase (pIVPRO, 10.88 kb) or mutant green fluorescent protein with e
nhanced fluorescence properties (mut3GFP, pIVET-GFP, 5.48 kb) as repor
ter gene systems. The proposed strategies can be adapted for use in mo
st Gram-negative bacteria.