Screening of target-specific stress-responsive genes for the development of cell-based biosensors using a DNA microarray.
In this study, we describe a straightforward strategy to develop whole cell-based biosensors using fusions of the bacterial bioluminescence genes and the promoters from chemically responsive genes within Escherichia coli, in which chemical target-responsive genes were screened by using the information of gene expression data obtained from DNA microarray analysis. Paraquat was used as a model chemical to trigger gene expression changes of E. coli and to show the DNA microarray-assisted development of whole cell-based biosensors. Gene expression data from the DNA microarray were obtained by time course analysis (10, 30, and 60 min) after exposure to paraquat. After clustering gene expression data obtained by time course analysis, a group of highly expressed genes over the all time courses could be classified. Within this group, three genes expressed highly for overall time points were selected and promoters of these genes were used as fusion partners with reporter genes, lux CDABE, to construct whole cell-based biosensors. The constructed biosensors recognized the presence of model inducer, paraquat, and structural analogue chemicals of paraquat with a high specificity, and the results were reconfirmed by using DNA microarray experiments for those structural analogues. This strategy to develop whole cell-based biosensors assisted by DNA microarray information should be useful in general for constructing chemical-specific or stress-specific biosensors with a high-throughput manner.
Kim
Byoung Chan
BC
Advanced Environmental Monitoring Research Center (ADEMRC), Gwangju Institute of Science and Technology (GIST), 1, Oryoung-dong, Puk-gu, Gwangju 500-712, Republic of Korea.
Youn
Chul Hee
CH
Ahn
Joo-Myung
JM
Gu
Man Bock
MB
D015374:Biosensing Techniques / Q000379:methods*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods*; D005798:Genes, Bacterial; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008164:Luminescent Proteins / Q000032:analysis; D016685:Mitomycin / Q000494:pharmacology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010269:Paraquat / Q000494:pharmacology
D008164:Luminescent Proteins; D016685:Mitomycin; D006861:Hydrogen Peroxide; D010269:Paraquat