Luminescent bacterial sensor for cadmium and lead.
A sensor plasmid was constructed by inserting the regulation unit from the cadA determinant of plasmid pI258 to control the expression of firefly luciferase. The resulting sensor plasmid pTOO24 is capable of replicating in Gram-positive and Gram-negative bacteria. The expression of the reporter gene as a function of added extracellular heavy metals was studied in Staphylococcus aureus strain RN4220 and Bacillus subtilis strain BR151. Strain RN4220(pTOO24) mainly responded to cadmium, lead and antimony, the lowest detectable concentrations being 10 nM, 33 nM and 1 nM respectively. Strain BR151(pTOO24) responded to cadmium, antimony, zinc and tin at concentrations starting from 3.3 nM, 33 nM, 1 microM, and 100 microM, respectively. The luminescence ratios between induced and uninduced cells, the induction coefficients, of strains RN4220(pTOO24) and BR151(pTOO24) were 23-50 and about 5, respectively. These results were obtained with only 2-3 h incubation times. Freeze-drying of the sensor strains had only moderate effects on the performance with respect to sensitivity or induction coefficients.
Tauriainen
S
S
University of Turku, Department of Biotechnology, Finland.
Karp
M
M
Chang
W
W
Virta
M
M
D000818:Animals; D001412:Bacillus subtilis / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D002104:Cadmium / Q000032:analysis* / Q000494:pharmacology; D001517:Coleoptera / Q000201:enzymology / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D005612:Freeze Drying; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D017930:Genes, Reporter / Q000187:drug effects; D007854:Lead / Q000032:analysis* / Q000494:pharmacology; D008156:Luciferases / Q000235:genetics; D008163:Luminescent Measurements; D010957:Plasmids / Q000235:genetics; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics
D017931:DNA Primers; D002104:Cadmium; D007854:Lead; D008156:Luciferases