Firefly luciferase as a reporter enzyme for measuring gene expression in vegetative and symbiotic Rhizobium meliloti and other gram-negative bacteria.
A DNA segment carrying a cDNA copy of the luciferase gene (luc) of the North American firefly Photinus pyralis, fused to the lambda PR promoter and expressed in Escherichia coli [de Wet et al., Proc. Natl. Acad. Sci. USA 82 (1985) 7870-7873], was inserted into a broad-host-range plasmid vector and established in a variety of Gram-negative bacteria. Luciferase activity, expressed from the lambda PR promoter, was detected in both intact cells and extracts prepared from cells of strains of Rhizobium meliloti, R. phaseoli, R. fredii, Pseudomonas aeruginosa, Agrobacterium tumefaciens, Acinetobacter calcoaceticus and Azotobacter vinelandii. The highest levels of activity, determined by measurements of both intact cells and extracts, were observed for P. aeruginosa and the three species of Rhizobium examined. Expression of luciferase activity also was relatively high in R. meliloti bacteroids of mature alfalfa nodules. This activity was readily detectable in intact nodules using x-ray film or in extracts prepared from purified bacteroids.
Palomares
A J
AJ
Department of Biology, University of California, San Diego, La Jolla 92093.
DeLuca
M A
MA
Helinski
D R
DR
D000818:Animals; D001517:Coleoptera / Q000201:enzymology; D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000096:biosynthesis / Q000302:isolation & purification; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006090:Gram-Negative Bacteria / Q000235:genetics*; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics*; D000455:Medicago sativa / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012231:Rhizobium / Q000235:genetics*; D013559:Symbiosis
D004269:DNA, Bacterial; D008156:Luciferases