Screening of target-specific stress-responsive genes for the development of cell-based biosensors using a DNA microarray

Byoung Chan Kim, Chul Hee Youn, Joo Myung Ahn, Man Bock Gu

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

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 bywith DNA microarray information should be useful in general for constructing chemical-specific or stress-specific biosensors with a high-throughput manner.

Original languageEnglish
Pages (from-to)8020-8026
Number of pages7
JournalAnalytical Chemistry
Volume77
Issue number24
DOIs
Publication statusPublished - 2005 Dec 15

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Microarrays
Biosensors
Screening
Genes
Paraquat
DNA
Gene expression
Escherichia coli
Fusion reactions
Bioluminescence
Throughput
Experiments

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Screening of target-specific stress-responsive genes for the development of cell-based biosensors using a DNA microarray. / Kim, Byoung Chan; Youn, Chul Hee; Ahn, Joo Myung; Gu, Man Bock.

In: Analytical Chemistry, Vol. 77, No. 24, 15.12.2005, p. 8020-8026.

Research output: Contribution to journalArticle

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