A cell array biosensor for environmental toxicity analysis.
In this study, a cell-based array technology that uses recombinant bioluminescent bacteria to detect and classify environmental toxicity has been implemented to develop two biosensor arrays, i.e., a chip and a plate array. Twenty recombinant bioluminescent bacteria, having different promoters fused with the bacterial lux genes, were immobilized within LB-agar. About 2 microl of the cell-agar mixture was deposited into the wells of either a cell chip or a 384-well plate. The bioluminescence (BL) from the cell arrays was measured with the use of highly sensitive cooled CCD camera that measured the bioluminescent signal from the immobilized cells and then quantified the pixel density using image analysis software. The responses from the cell arrays were characterized using three chemicals that cause either superoxide damage (paraquat), DNA damage (mitomycin C) or protein/membrane damage (salicylic acid). The responses were found to be dependent upon the promoter fused upstream of the lux operon within each strain. Therefore, a sample's toxicity can be analyzed and classified through the changes in the BL expression from each well. Moreover, a time of only 2 h was needed for analysis, making either of these arrays a fast, portable and economical high-throughput biosensor system for detecting environmental toxicities.
Lee
Jin Hyung
JH
National Research Laboratory on Environmental Biotechnology, Gwangju Institute of Science and Technology (GIST), 1 Oryong-dong, Buk-gu, Gwangju 500-712, Republic of Korea.
Mitchell
Robert J
RJ
Kim
Byoung Chan
BC
Cullen
David C
DC
Gu
Man Bock
MB
D029681:Arabidopsis Proteins / Q000378:metabolism; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D004784:Environmental Monitoring / Q000295:instrumentation* / Q000379:methods; D004785:Environmental Pollutants / Q000009:adverse effects*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D046228:Microarray Analysis / Q000295:instrumentation* / Q000379:methods; D018675:Toxicity Tests / Q000295:instrumentation* / Q000379:methods; D014157:Transcription Factors / Q000378:metabolism
D029681:Arabidopsis Proteins; D004785:Environmental Pollutants; D008164:Luminescent Proteins; C504848:Lux Arrhythmo protein, Arabidopsis; D014157:Transcription Factors