• Journal of Microbiology and Biotechnology
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• v.12 no.3
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• pp.417-422
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• 2002

Gene fusions were constructed by the transcriptional fusion of the dnaK promoter of pseudomonas sp. DJ-12 or E. coli to the lux or luc marker gene. The dnaKp-DJ::luxCDABE bioluminescent fusion in the biosensor using the Pseudomonas sp. DJ-12 dnaK promoter exhibited about 5-fold more extensive response to ethanol than that of dnaKp-EC::luxCDABE. The bioluminescent response of the dnaK-DJ::luc fusion to ethanol was much weaker than those of the other fusions. The biosensor harboring the dnaKp-DJ::luCDABE fusion was examined for its bioluminescence production based on exposure to aromatic compounds, such as biphenyl, 4-chlorobiphenyl (4CB), 4-hydroxybenzoate (4HBA), and catechol. In particular, the bioluminescence produced by the dnaKp-DJ::luxCDABE fusion was most sensitive to 1 mM biphenyl and 4CB when exposed for 80 min, and the responses were also very strong to other aromatics. Therefore, the biosensor bearing the dnaKp-DJ::luxCDABE fusion would appear to be the most useful for the detection of aromatics and other pollutants.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.2
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• pp.101-105
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• 2003

Recombinant bioluminescent bacteria were used to monitor and classify the to xicity of azo dyes. Two constitutive bioluminescent bacteria, Photobacterium phosphoreum and Es-Cherichia coli, E, coli GC2 (lac::luxCOABE), were used to detect the cellular toxicity of the azo dyes. In addition, four stress-inducible bioluminestent E. coli, DPD2794 (recA::luxCDABE), a DNA damage Sensitive strain; DPD2540 (fabA::luxCDABE), a membrane damage sensitive strain; DPD2511 (katG::luxCDABE), an oxidative damage sensitive strain; and TV1061 (grpE::luxCDABE), a protein damage sensitive strain, were used to provide information about the type of toxicity caused by crystal violet, the most toxic dye of the 16 azo dyes tested. These results suggest that azo dyes result in serious cellular toxicity in bacteria, and that toxicity monitoring and classific ation of some azo dyes, In the field, may be possible using these recombinant bioluminescent bacteria.

• Journal of Microbiology and Biotechnology
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• v.15 no.1
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• pp.48-54
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• 2005

A comparison of promoter fusions with the luxCDABE genes from Vibrio fischeri and Photorhabdus luminescens was made using promoters from several genes (katG, sodA, and pqi-5) of E. coli that are responsive to oxidative damage. The respective characteristics, such as the basal and maximum bioluminescence and the relative bioluminescence, were compared. E. coli strains carrying fusions of the promoters to P. luminescens lux showed higher basal and maximally induced bioluminescent levels than strains carrying the same promoter fused to the luxCDABE genes from V. fischeri. The sensitivities between the strains were similar, regardless of the luciferase used, but lower response ratios were seen from strains harboring the P. luminescens lux fusions. Furthermore, using the two katG::lux fusion strains, the bioluminescence from the P. luminescens lux fusion strain, DK1, was stable after reaching a maximum, while that of strain DPD2511 decreased very rapidly due to substrate limitation.

• Journal of Microbiology and Biotechnology
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• v.13 no.1
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• pp.99-103
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• 2003

An Escherichia coli strain containing the recA promoter that fused to the luxCDABE operon originating from Photorhabdus luminescens was shown to respond sensitively to genotoxic stresses. Two different recombinant bacteria, one (DPDI 657) harboring a plasmid with the recA promoter that fused to the luxCDABE operon, and the other (DPD1710) containing a chromosomally-integrated recA promoter that fused with luxCDABE, were compared and it was found that the sensitivity of 'the two strains was significantly different in terms of their bioluminescent level, response time, and the minimum detectable concentration of a chemical causing DNA damaging stress. DPDI 710, with a chromosomally-integrated single copy, generally led to lower basal luminescence levels, faster responses, increased response ratios, and an enhanced sensitivity to mutagens, when compared to DPD 1657 with a multi-copy plasmid.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.6
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• pp.395-399
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• 2000

Five different freeze-dried recombinant bioluminescent bacteria were used for the detection of cellular stresses caused by endocrine disrupting chemicals. These strains were DPD2794 (recA::luxCDABE), which is sensitive to DNA damage, DPD2540 (fabA::luxCDABE), sensitive to cellular membrane damage, DPD2511 (katG::luxCDABE), sensitive to oxidative damage, and TV1061 (grpE::luxCDABE), sensitive to protein damage. GC2, which emits bioluminescence constitutively, was also used in this study. The toxicity of several chemicals was measured using GC2. Damage caused by known endocrine disrupting chemicals, such as nonyl phenol, bisphenol A, and styrene, was detected and classified according to toxicity mode, while others, such as phathalate and DDT, were not detected with the bacteria. These results suggest that endocrine disrupting chemicals are toxic in bacteria, and do not act via an estrogenic effect, and that toxicity monitoring and classification of some endocrine disrupting chemicals may be possible in the field using these freeze-dried recombinant bioluminescent bacteria.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.187-190
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• 2000

Effects of gamma ionizing radiation on recombinant Escherichia coli cells containing stress promoters, recA, fabA, grpE, or katG, fused to luxCDABE originated from Vibrio fischeri were characterized by monitoring transcriptional responses reflected by bioluminescent output. Quantification of gamma-ray intensity may be possible using the recA and fabA promoter fusion since a linear enhancement of bioluminescence emission with increasing gamma-ray intensity was observed. Other strains sensitive to either oxidative stress (DPD2511, katG::luxCDABE) or protein-damaging stress (TV1061, grpE::luxCDABE) were also irradiated by gamma-rays, and resulted in no noticeable bioluminescent output while DPD2794 with recA promoter and DPD2540 with fabA promoter irradiated by the same intensities of gamma-rays gave a significant bioluminescent output. This indicates that the main stresses in the recombinant bacteria caused by ionizing radiation are DNA and membrane-damage, not protein- or oxidative-damage. In addition, in this study, to investigate the relationship between the radiation dose rate and bacterial responses, two recombinant Escherichia coli strains, DPD2794 and GC2, containing lac promoter fused to luxCDABE originating from Photorhapdus luminescences, were used for detecting DNA damage and cellular toxicity under various radiation dose rates. Throughout this study, it was found that these bacteria showed quantitative stress responses to DNA damage and general toxicity caused by gamma rays, depending on the radiation dose rates, indicating that the bacterial stress responses and general toxicity were seriously influenced according to radiation dose rates.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.6
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• pp.516-521
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• 2006

The use of gltA gene, as a new biomarker for environmental stress biomonitoring, was investigated because of its key position as the first enzyme of the tricarboxylic acid (TCA) cycle. A recombinant bioluminescent Escherichia coli strain, EBJM2, was constructed using a plasmid carrying the citrate synthase (gltA) promoter transcribing the Photorhabdus luminescens IuxCDABE genes (gltA::luxCDABE). The responses from this strain were studied with five different classes of toxicants: DNA damage chemicals, phenolics, oxidative-stress chemicals, PAHs, and organic solvents. EBJM2 responded strongly to DNA damage chemicals, such as mitomycin C (MMC) and methyl-nitro-nitrosoguanidine (MNNG) and nalidixic acid with the strongest responses. In contrast, tests with several compounds from the other four classes of toxicants gave no significant response. Therefore, EBJM2 was found to be sensitive to DNA damage chemicals.

• Journal of Microbiology and Biotechnology
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• v.7 no.5
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• pp.352-355
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• 1997

The promoter region of the pupA gene of Pseudomonas putida WCS358 was fused with the structural genes for bioluminescence (luxCDABE) from Vibrio fischeri, and the resulting fusion plasmid harbored by the WCS358 host. The pup-lux fusion gene was then used for quantitative analysis of the iron-dependence of pupA promoter activity. Factors affecting bioluminescence produced by the pup-lux bioreporter were found to be cell activity, iron-chelator concentrations, Fe(III) concentrations, and nutrient components. Light production rates of the pup-lux bioreporter were inversely dependent upon iron molecules when $FeCl_3$ concentrations were between $10^{-2}$ and 1 ${\mu}M$ in nutrient-poor minimal media, and between 0.1 and 10 mM in nutrient-rich complex media.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.688-688
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• 2003

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.645-646
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• 2003