Screening of Target-Specific Stress-Responsive Genes for the Development of Cell-Based Biosensors Using a DNA Microarray

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• 作者：Byoung Chan Kim ; Chul Hee Youn ; Joo-Myung Ahn ; and Man Bock Gu
• 刊名：Analytical Chemistry
• 出版年：2005
• 出版时间：December 15, 2005
• 年：2005
• 卷：77
• 期：24
• 页码：8020 - 8026
• 全文大小：334K
• 年卷期：v.77,no.24(December 15, 2005)
• ISSN：1520-6882

文摘

In this study, we describe a straightforward strategy todevelop whole cell-based biosensors using fusions of thebacterial bioluminescence genes and the promoters fromchemically responsive genes within Escherichia coli, inwhich chemical target-responsive genes were screened byusing the information of gene expression data obtainedfrom DNA microarray analysis. Paraquat was used as amodel chemical to trigger gene expression changes of E.coli and to show the DNA microarray-assisted development of whole cell-based biosensors. Gene expressiondata from the DNA microarray were obtained by timecourse analysis (10, 30, and 60 min) after exposure toparaquat. After clustering gene expression data obtainedby time course analysis, a group of highly expressed genesover the all time courses could be classified. Within thisgroup, three genes expressed highly for overall time pointswere selected and promoters of these genes were usedas fusion partners with reporter genes, lux CDABE, toconstruct whole cell-based biosensors. The constructedbiosensors recognized the presence of model inducer,paraquat, and structural analogue chemicals of paraquatwith a high specificity, and the results were reconfirmedby using DNA microarray experiments for those structuralanalogues. This strategy to develop whole cell-basedbiosensors assisted bywith DNA microarray informationshould be useful in general for constructing chemical-specific or stress-specific biosensors with a high-throughput manner.