• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.466-469
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• 2004

The recombinant bioluminescent Escherichia coli strains, DPD2511 and TV 1061 containing the katG and grpE promoters, respectively, from Vibrio fischeri fused to luxCDABE, were used to detect the adaptive and repair responses to oxidative damage caused by hydrogen peroxide $(H_2O_2)$, and protein damage due to phenol. The response ratio, represented as the bioluminescence induced in subsequent inductions of DPD2511 and TV1061 with the mother cells previously induced by each chemical, i.e., $H_2O_2$ and phenol during the previous induction stage, decreased suddenly compared with the ratio of the control culture of each strain, meaning there is a possible adaptive response to stress caused by chemicals. Protein damage due to phenol was completely repaired by the second culturing after the initial induction, as was oxidative damage caused by $H_2O_2$ which was also rapidly repaired, as detected by the recovery of bioluminescence level. This result suggests that E. coli promptly adapt and repair oxidative and protein damage by $H_2O_2$ and phenol completely.

• Journal of Soil and Groundwater Environment
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• v.18 no.4
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• pp.32-39
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• 2013

This research focused on the effects of heavy metals on the biomonitoring activity of genetically engineered bioluminescent bacteria, Pseudomonas putida mt-2 KG1206. KG1206 was exposed to single or binary mixtures of different heavy metals as well as soils contaminated with heavy metals. In case of single exposure with different inducer pollutant, the toxicity order was as followings : As(III) > Cd, As(V) >> Cu, Cr(VI). The toxic effects of the binary mixtures was compared to the expected effect based on a simple theory of probabilities. The interactive effects were mostly additive, while in few cases antgonistic and synergistic mode of action was observed for some concentration combinations. No considerable correlation was found between the total metal contents in soils and the bioluminescence activity of each sample. However, statistically significant differences (p = 0.0102) were observed between two groups, classified based on arsenite contamination. These results demonstrate the usage of recombinant bioluminescent microorganism in biomonitoring and the complex interactive effects of metals.

• Biotechnology and Bioprocess Engineering:BBE
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• v.2 no.2
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• pp.141-146
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• 1997

A new sensing system based on the immobilization of luminescent batcteria, Photobacterium phosphoreum, was proposed for continuous real-time monitoring of polluants. The response curves demonstrate that Photobacterium phosphoreum immobilized on the strontium alginate was very sensitive to seven reference chemicals used. The significant inhibitory concentrations for bioluminescence emission were 5 ppm for Pb(NO3)2, NiCl2, CdCl2, 50 ppm for NaAsO2, 0.1ppm for HgCl2, 0.5ppm for pentachlorophenol and less than 5ppm for SDS, respectively. The alginate mixed-cells (AMC) retained their luminescence during experimental period (29 days) under storage condition of -8$0^{\circ}C$. The variables affecting performance of continuous flow through monitoring (CFTM) were optimized in order to ensure stability and efficiency. The flow through cell with strontium-alginate immobilized luminescent bacteria was tested with salicylate and 4-nitrophenol and a rapid response of luminescence was recorded by time drive mode in bioluminescence spectrometer after exposure to both toxicants.

• Journal of Biomedical Engineering Research
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• v.32 no.2
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• pp.85-92
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• 2011

Bioluminescence imaging (BLI) is the most sensitive animal imaging technique for molecular imaging research. Generally, highly sensitive CCD is used to detect an optical probe introduced in a living mouse. However, in many cases, the light signal emitted from a probe is too small to detect because it is scattered and attenuated by the tissue prior to being detected. The problem is that scattering and attenuation not only inhibit accurate measurement but also make image quality down. Thus we introduced a new method to reduce noise by using property of CCD and method to improve image quality of bioluminescence image by using two steps Gaussian blurring.

• Journal of Korean Neurosurgical Society
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• v.48 no.5
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• pp.391-398
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• 2010

Objective : This study was designed to validate the cell trafficking efficiency of the in vivo bioluminescence image (BLI) study in the setting of transplantation of the luciferase expressing bone marrow-derived mesenchymal stem cells (BMSC), which were delivered at each different time after transient middle cerebral artery occlusion (MCAO) in a mouse model. Methods : Transplanting donor BMSC were prepared by primary cell culture from transgenic mouse expressing luciferase (LUC). Transient focal infarcts were induced in 4-6-week-old male nude mice. The experiment mice were divided into five groups by the time of MSC transplantation : 1) sham-operation group, 2) 2-h group, 3) 1-day group, 4) 3-day group, and 5) 1-week group. BLI for detection of spatial distribution of transplanted MSC was performed by detecting emitted photons. Migration of the transplanted cells to the infarcted area was confirmed by histological examinations. Differences between groups were evaluated by paired t-test. Results : A focal spot of bioluminescence was observed at the injection site on the next day after transplantation by Signal intensity of bioluminescence. After 4 weeks, the mean signal intensities of 2-h, 1-day, 3-day, and 1-week group were $2.6{\times}10^7{\pm}7.4{\times}10^6$. $6.1{\times}10^6{\pm}1.2{\times}10^6$, $1.7{\times}10^6{\pm}4.4{\times}10^5$, and $8.9{\times}10^6{\pm}9.5{\times}10^5$, respectively. The 2-h group showed significantly higher signal intensity (p<0.01). The engrafted BMSC showed around the infarct border zones on immunohistochemical examination. The counts of LUC-positive cells revealed the highest number in the 2-h group, in agreement with the results of BLI experiments (p<0.01). Conclusion : In this study, the results suggested that the transplanted BMSC migrated to the infarct border zone in BLI study and the higher signal intensity of LUC-positive cells seen in 2 hrs after MSC transplantation in MCAO mouse model. In addition, noninvasive imaging in real time is an ideal method for tracking stem cell transplantation. This method can be widely applied to various research fields of cell transplantation therapy.

• Journal of Life Science
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• v.20 no.11
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• pp.1738-1741
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• 2010

Caliper measurements of tumor volume have been widely used in the assessment of tumors in animal models. However, experiments based on caliper data have resulted in unreliable estimates of tumor growth, due to necrotic areas of tumor mass. To overcome this systematic bias, we engineered a new luciferase-expressing rat prostate cancer cell line (MLL-Luc) that produces bioluminescence from viable cancer cells. MLL-Luc cells showed a strong correlation between bioluminescence intensity and cell number ($R^2$=0.99) and also accurately quantified tumor growth, with reduced bioluminescence signals caused by necrotic cells in a subcutaneous MLL-Luc xenograft model. The accurate quantification of tumor growth with bioluminescence imaging (BLI) was confirmed by a better antitumor effect of combination chemotherapy, compared to that based on caliper measurements with a correlation between the bioluminescence signal and tumor volume ($R^2$=0.84). These data suggest that bioluminescent MLL xenografts are a powerful and quantitative tool for monitoring tumor growth and are useful in evaluating the efficacy of anticancer drugs, with less systematic bias.

• Bulletin of Food Technology
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• v.11 no.3
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• pp.129-138
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• 1998

• 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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• 2000.11a
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• pp.212-215
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• 2000

Recombinant bioluminescent bacterial strains that use specific promoters fused to the bioluminescence genes (lux genes) have been applied in environmental monitoring. Advantages of using recombinant bioluminescent bacteria as blosensing cells include rapid responses, low costs, and improved reproducibility. In this study, a recombinant Escherichia coli, GC2, containing a lac::luxCDABE fusion immobilized with solid agar media and glass beads was used to estimate the effect of this soil flushing technique. This bacterium constitutively emits light under normal conditions (no toxic chemicals). When growth and metabolism of these bioluminescent bacteria is inhibited by their exposure to toxic chemicals, the bioluminescence (BL) is reduced. A biosurfactant, rhamnolipids, was used to extract phenanthrene from the soil after flushing.

• Journal of Microbiology
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• v.38 no.2
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• pp.74-79
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• 2000

For the demonstration of a novel riboflavin kinase assay method based on the bacterial bioluminescence, partially purified riboflavin kinase was prepared from bovine liver through ammonium sulfate precipitation and DEAE-cellulose ion exchange chromatography. Using bacterial luciferase from Photobacterium phosphoreum and the dithionite reduction method, and easy, safe, and fast assay method was established. The optimal temperature, pH, Km values form riboflavin and ATP of boving liver riboflavin kinase determined with this luminescence method were 35$^{\circ}C$, pH 7, 15.3${\mu}$M and 8.3.${\mu}$M, respectively. The detection limit of FMN produced by riboflavin kinase was in the range of 200 pM to 4${\mu}$M which is comparable to the HPLC-fluorescence detection method, while the detection time for each assay was less than 15 sec compared to the HPLC method which requeires at least 10 min for completion.