Electrochemical mutagen screening using microbial chip.

Electrochemical microbial chip for mutagen screening were microfabricated and characterized by scanning electrochemical microscopy (SECM). Salmonella typhimurium TA1535 with a plasmid pSK1002 carrying a umuC'-'lacZ fusion gene was used for the whole cell mutagen sensor. The TA1535/pSK1002 cells were exposed to mutagen solutions containing 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamido (AF-2), mitomycin C (MMC) or 2-aminoanthracene (2-AA) and embedded in a microcavity (5nl) on a glass substrate using collagen gel. The beta-galactosidase expression on the microbial chip was electrochemically monitored using p-aminophenyl-beta-d-galactopyranoside (PAPG) as the enzymatic substrate. This system has several advantages compared with the conventional umu test: drastic reduction of the sample volume, less time-consuming for beta-galactosidase detection (free from substrate reaction time) and lower detection limit for the three mutagens (AF-2, MMC, 2-AA). Finally, a multi-sample assay was carried out using the microbial array chip with four microcavities.

[1]  A. Bard,et al.  Chemical Imaging of Surfaces with the Scanning Electrochemical Microscope , 1991, Science.

[2]  Shimshon Belkin,et al.  Genotoxicity monitoring using a 2D-spectroscopic GFP whole cell biosensing system , 2003 .

[3]  B. Ames,et al.  An improved bacterial test system for the detection and classification of mutagens and carcinogens. , 1973, Proceedings of the National Academy of Sciences of the United States of America.

[4]  H. Obana,et al.  Suppressive effects of coffee on the SOS responses induced by UV and chemical mutagens. , 1986, Mutation research.

[5]  Teruyuki Nagamune,et al.  Construction of green fluorescent protein reporter genes for genotoxicity test (SOS/umu-test) and improvement of mutagen-sensitivity. , 2001, Journal of bioscience and bioengineering.

[6]  J. Kulys,et al.  Electrochemical oxidation of catechol and p-aminophenol esters in the presence of hydrolases , 1980 .

[7]  A. Bard,et al.  Scanning Electrochemical and Tunneling Ultramicroelectrode Microscope for High-Resolution Examination of Electrode Surfaces in Solution , 1986 .

[8]  M. Hofnung,et al.  SOS chromotest, a direct assay of induction of an SOS function in Escherichia coli K-12 to measure genotoxicity. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[9]  J Rishpon,et al.  Online and in situ monitoring of environmental pollutants: electrochemical biosensing of cadmium. , 2000, Environmental microbiology.

[10]  N. Kado,et al.  A simple modification of the Salmonella liquid-incubation assay. Increased sensitivity for detecting mutagens in human urine. , 1983, Mutation research.

[11]  G. Barrett,et al.  Genetically engineered whole-cell sensing systems: coupling biological recognition with reporter genes. , 2000, Chemical reviews.

[12]  H. Shinagawa,et al.  Mutational specificity of the umuC mediated mutagenesis in Eschericha coli. , 1982, Biochimie.

[13]  T. Shimada,et al.  SOS-inducing activity of chemical carcinogens and mutagens in Salmonella typhimurium TA1535/pSK1002: examination with 151 chemicals. , 1987, Mutation research.

[14]  Bioluminescence detection system of mutagen using firefly luciferase genes introduced in Escherichia coli lysogenic strain , 1992 .

[15]  J. Rishpon,et al.  Electrochemical detection of protein-protein interactions using a yeast two hybrid: 17-beta-estradiol as a model. , 2003, Analytical biochemistry.

[16]  Reuven Babai,et al.  On-line and in situ biosensors for monitoring environmental pollution. , 2003, Biotechnology advances.

[17]  B. Ames,et al.  Revised methods for the Salmonella mutagenicity test. , 1983, Mutation research.

[18]  Makino Kozo,et al.  Cloning and characterization of the umu operon responsible for inducible mutagenesis in Escherichia coli , 1983 .

[19]  M. Plewa,et al.  Mutagenicity of atrazine: a maize-microbe bioassay. , 1976, Mutation research.

[20]  R D Schmid,et al.  Reporter gene bioassays in environmental analysis , 2000, Fresenius' journal of analytical chemistry.

[21]  Matsuhiko Nishizawa,et al.  Scanning electrochemical microscopy-based drug sensitivity test for a cell culture integrated in silicon microstructures. , 2003, Analytical chemistry.

[22]  M. Kitano,et al.  Use of a high‐throughput umu‐microplate test system for rapid detection of genotoxicity produced by mutagenic carcinogens and airborne particulate matter , 2004, Environmental and molecular mutagenesis.

[23]  H. Shinagawa,et al.  Evaluation of the new system (umu-test) for the detection of environmental mutagens and carcinogens. , 1985, Mutation research.

[24]  Hirofumi Tani,et al.  Chip-based bioassay using bacterial sensor strains immobilized in three-dimensional microfluidic network. , 2004, Analytical chemistry.

[25]  David R Walt,et al.  Living bacterial cell array for genotoxin monitoring. , 2004, Analytical chemistry.

[26]  G. Reifferscheid,et al.  Increase of sensitivity and validity of the SOS/umu-test after replacement of the beta-galactosidase reporter gene with luciferase. , 1997, Mutation research.

[27]  D. Mount,et al.  The SOS regulatory system of Escherichia coli , 1982, Cell.

[28]  H. Maki.  Origins of spontaneous mutations: specificity and directionality of base-substitution, frameshift, and sequence-substitution mutageneses. , 2002, Annual review of genetics.

[29]  I Karube,et al.  Microbial sensor for preliminary screening of mutagens utilizing a phage induction test. , 1989, Analytical chemistry.

[30]  T. Horii,et al.  Structural analysis of the umu operon required for inducible mutagenesis in Escherichia coli. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[31]  G. Węgrzyn,et al.  Detection of mutagenic pollution of natural environment using microbiological assays , 2003, Journal of applied microbiology.

[32]  C. Wijayawardhana,et al.  Monitoring β-galactosidase activity by means of scanning electrochemical microscopy , 2004 .

[33]  M. Henry-Amar,et al.  Biomonitoring of urine mutagenicity with the Ames test: improvement of the extraction/concentration method. , 2002, Mutation research.

[34]  H. Shiku,et al.  Fabrication of microbial chip using collagen gel microstructure. , 2003, Lab on a chip.

[35]  J Rishpon,et al.  On-line monitoring of gene expression. , 1999, Microbiology.

[36]  R. Larossa,et al.  Improved bacterial SOS promoter∷lux fusions for genotoxicity detection. , 2000, Mutation research.

[37]  H. Shiku,et al.  On-chip electrochemical measurement of beta-galactosidase expression using a microbial chip. , 2004, Chemical communications.

[38]  M. Miyazawa,et al.  Antimutagenic activity of phenylpropanoids from clove (Syzygium aromaticum). , 2003, Journal of agricultural and food chemistry.

[39]  B. Ames,et al.  Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test. , 1975, Mutation research.

[40]  David Juncker,et al.  Simultaneous detection of C-reactive protein and other cardiac markers in human plasma using micromosaic immunoassays and self-regulating microfluidic networks. , 2004, Biosensors & bioelectronics.

[41]  C. Janion,et al.  Historical Aspects , 1957 .