Abstract
The bioassay LUX-FLUORO test was developed for the rapid detection and quantification of environmental pollutants with genotoxic and/or cytotoxic effects. This bacterial test system uses two different reporter genes whose gene products and their reactions, respectively, can be measured easily and simultaneously by optical methods. Genotoxicity is measured by the increase of bioluminescence in genetically modified bacteria which carry a plasmid with a complete lux operon for the enzyme luciferase from the marine photobacterium P. leiognathi under the control of a DNA-damage dependent so-called SOS promoter. If the desoxyribonucleic acid (DNA) in these bacteria is damaged by a genotoxic chemical, the SOS promoter is turned on and the lux operon is expressed. The newly synthesized luciferase reacts with its substrate thereby producing bioluminescence in a damage-proportional manner. In the second part of the system, genetically modified bacteria carry the gfp gene for the green fluorescent protein from the jellyfish A. victoria downstream from a constitutively expressed promoter. These bacteria are fluorescent under normal conditions. If their cellular metabolism is disturbed by the action of cytotoxic chemicals the fluorescence decreases in a dose-proportional manner. The combined LUX-FLUORO test can be used for the biological assessment of the geno- and cytotoxicity of a wide variety of organic and inorganic chemicals including complex mixtures in different matrices.
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References
Albertini RJ (2001) HPRT mutations in humans: biomarkers for mechanistic studies. Mutat Res 489:1–16
Ames BN, Lee FD, Durston WE (1973) An improved bacterial test system for the detection and classification of mutagens and carcinogens. Proc Natl Acad Sci USA 70:782–786
Arfsten DP, Davenport R, Schaeffer DJ (1994) Reversion of bioluminescent bacteria (Mutatox™) to their luminescent state upon exposure to organic compounds, munitions, and metal salts. Biomed Environ Sci 7:144–149
Baldwin TO, Christopher JA, Raushel FM, Sinclair JF, Ziegler MM, Fisher AJ, Rayment I (1995) Structure of bacterial luciferase. Current Opinion in Structural Biology 5:798–809
Baumstark-Khan C, Rode A, Rettberg P, Horneck G (2001) Application of the lux-fluoro-test as bioassay for combined genotoxicity and cytotoxicity measurements by means of recombinant Salmonella typhimurium TA1535 cells. Analyt Chim Acta 437:23–30
Bianchi V, Celotti L, Lanfranchi G, Majore F, Marin G, Montaldi A, Sponza G, Tamino G, Nenier P, Zantedechi A, Levis AG (1983) Genetic effects of chromium compounds. Mutat Res 117:279–300
Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC (1994) Green fluorescent protein as a marker for gene expression. Science 263:802–805
Crameri A, Whitehorn EA, Tate E, Stemmer WP (1996) Improved green fluorescent protein by molecular evolution using DNA shuffling. Nat Biotechnol 14:315–319
D’Souza SF (2001) Microbial biosensors. Biosens Bioelectron 16:337–353
Enzmann H, Bomhard E, Iatropoulos M, Ahr HJ, Schlueter G, William GM (1998) Short-and intermediate-term carcinogenicity testing — a review. Part 1: The prototypes mouse skin tumour assay and rat liver focus assay. Food Chem Toxicol 36:979–995
Friedberg EC (1985) DNA repair. Freeman and Company, New York
Horneck G, Ptitsyn LR, Rettberg P, Komova O, Kozubek S, Krasavin EA (1998) Recombinant Escherichia coli cells as biodetector system for genotoxins. In: Hock B, Barceló D, Cammann K, Hansen P-D, Turner APF (eds) Biosensors for environmental diagnostics. Taubner-Reihe UMWELT, Stuttgart, pp 215–232
ICH (1997) Harmonized Tripartite Guideline, 2SB, Genotoxicity: Standard Battery Tests, CPMP/ICH/ 174/95, http://www.ifpma.org/ich5s.html
Lantzsch H, Gebel T (1997) Genotoxicity of selected metal compounds in the SOS chromotest. Mutat Res 389:191–197
Mortelmans K, Zeiger E (2000) The Ames Salmonella/microsome mutagenicity assay. Mutat Res 455:29–60
Natarajan T, Balajee AS, Boei JJWA, Darroudi F, Dominguez I, Hande MP, Meijers M, Slijepcevic P, Vermeulen S, Xiao Y (1996) Mechanisms of induction of chromosomal aberrations and their detection by fluorescence in-situ hybridization. Mutat Res 372:247–258
Ptitsyn LR, Horneck G, Komova O, Kozubek S, Krasavin EA, Bonev M, Rettberg P (1997) A biosensor for environmental genotoxin screening based on an SOS lux assay in recombinant Escherichia coli cells. Appl Envir Microbiol 63:4377–4384
Quillardet P, Hofnung M (1993) The SOS chromotest: a review. Mutat Res 297:235–279
Rabbow E, Rettberg P, Baumstark-Khan C, Horneck G (2002) The SOS-LUX-and LAC-FLUORO-TEST for the quantification of genotoxic and/or cytotoxic effects of heavy metal salts. Analyt Chim Acta 456:31–39
Rettberg P, Baumstark-Khan C, Bandel K, Ptitsyn LR, Horneck G (1999) Microscale application of the SOS-LUX-TEST as biosensor for genotoxic agents. Anal Chim Acta 387:289–296
Rettberg P, Bandel K, Baumstark-Khan C, Horneck G (2001) Increased sensitivity of the SOS-LUX-test for the detection of hydrophobic genotoxic substances with Salmonella typhimurium TA1535 as host strain. Anal Chim Acta 426:167–173
White PA, Rasmussen JB (1996) SOS chromotest results in a broader context: empirical relationships between genotoxic potency, mutagenic potency, and carcinogenic potency. Environ Mol Mutagen 27:270–305
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P., R., Baumstark-Khan, C., Rabbow, E., Horneck, G. (2005). The LUX-FLUORO Test as a Rapid Bioassay for Environmental Pollutants. In: Lichtfouse, E., Schwarzbauer, J., Robert, D. (eds) Environmental Chemistry. Springer, Berlin, Heidelberg. https://doi.org/10.1007/3-540-26531-7_59
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DOI: https://doi.org/10.1007/3-540-26531-7_59
Publisher Name: Springer, Berlin, Heidelberg
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