• Biotechnology and Bioprocess Engineering:BBE
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• 제8권2호
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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.

• 한국데이터정보과학회:학술대회논문집
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• 한국데이터정보과학회 2006년도 PROCEEDINGS OF JOINT CONFERENCEOF KDISS AND KDAS
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• pp.121-129
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• 2006

Two color or cDNA microarrays are extensively used to study relative expression levels of thousands of genes simultaneously. 0かy two tissue samples can be hybridized on a single microarray slide. Thus, a microarray slide necessarily forms an incomplete block design with block size two when more than two tissue samples are under study. We also need to control for variability in gene expression values due to the two dyes. Thus, red and green dyes form the second blocking factor in addition to slides. General design problem for these microarray experiments is discussed in this paper. Designs for factorial cDNA microarrays are also discussed.

• Biomolecules & Therapeutics
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• 제1권1호
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• pp.58-64
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• 1993

DNA addicts induced by putative chemical related to carcinogenesis were detected and determined by $^{32}$P-Postlabelling assay after exposure of 4 compounds comprising two auto dyes (amaranth, new coccine) and two flavonoid compounds (rutin, quercetin) to ICR mouse. DNA was isolated from mouse liver and digested enzymatically to deoxyribonucleoside 3'-monophosphate. The postincubation of DNA digests with nuclease Pl before $^{32}$P-labelling enhanced the technique's sensitivity. Nuclease Pl cleaves deoxyribonucleoside 3'-mono-phosphates of normal nucleotides to deoxyrihonucleosides which do not serve as substrates for polynucleotide kinase, while most of addicts were found to be totally or partially resistant to the 3'-dephosphorylating action of nuclease Pl. The adducted deoxyribonucleoside 3'-monophosphate was converted to 5'-$^{32}$P-labelled deoxynucleoside 3',5'-bisphosphate by T4 polynucleotide kinase. The nucleotides were separated by anion-exchange thin layer chromatography(TLC) on polyethyleneimine cellulose by 4-dimensional or 2-dimensional TLC then detected by autoradiography. The results show that DNA addicts were detected in liver DNA of ICR mouse after administration of amaranth and quercetin by 2-dimensional and/or 4-dimensional TLC.

• 한국염색가공학회:학술대회논문집
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• 한국염색가공학회 2008년도 제39차 학술발표회
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• pp.133-134
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• 2008

A novel near infrared(NIR) absorbing dyes were synthesized by using bis-aldehyde formyl aromatic compounds and heteroaryl derivatives with the reactive methylene group. These dyes provided a range of the NIR wavelength region about 720 nm value. Also, the light absorbing properties of these dyes were investigated in our experiment results. In generally, these NIR colorants may be potential used for optical recording media, DNA sequencing probe and laser beam printings.

• Molecules and Cells
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• 제27권4호
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• pp.391-396
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• 2009

Image-based, high-content screening assays demand solutions for image segmentation and cellular compartment encoding to track critical events - for example those reported by GFP fusions within mitosis, signalling pathways and protein translocations. To meet this need, a series of nuclear/cytoplasmic discriminating probes have been developed: DRAQ5$^{TM}$ and CyTRAK Orange$^{TM}$. These are spectrally compatible with GFP reporters offering new solutions in imaging and cytometry. At their most fundamental they provide a convenient fluorescent emission signature which is spectrally separated from the commonly used reporter proteins (e.g. eGFP, YFP, mRFP) and fluorescent tags such as Alexafluor 488, fluorescein and Cy2. Additionally, they do not excite in the UV and thus avoid the complications of compound UV-autofluorescence in drug discovery whilst limiting the impact of background sample autofluorescence. They provide a convenient means of stoichiometrically labelling cell nuclei in live cells without the aid of DMSO and can equally be used for fixed cells. Further developments have permitted the simultaneous and differential labelling of both nuclear and cytoplasmic compartments in live and fixed cells to clearly render the precise location of cell boundaries which may be beneficial for quantitative expression measurements, cell-cell interactions and most recently compound in vitro toxicology testing.

• BioChip Journal
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• 제12권4호
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• pp.340-347
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• 2018

$F{\ddot{o}}rster$ resonance energy transfer (FRET) is extremely sensitive to the separation distance between the donor and the acceptor which is ideal for probing such biological phenomena. Also, FRET-based probes have been developing for detecting an unamplified, low-abundance of target DNA. Here we describe the development of FRET based DNA sensor based on an accumulated QD system for detecting KRAS G12D mutation which is the most common mutation in cancer. The accumulated QD system consists of the polystyrene beads which surface is modified with carboxyl modified QDs. The QDs are sandwich-hybridized with DNA of a capture probe, a reporter probe with Texas-red, and a target DNA by EDC-NHS coupling. Because the carboxyl modified QDs are located closely to each other in the accumulated QDs, these neighboring QDs are enough to transfer the energy to the acceptor dyes. Therefore the FRET factor in the bead system is enhancing by the additional increase of 29.2% as compared to that in a single QD system. These results suggest that the accumulated nanobead probe with conjugated QDs can be used as ultrasensitive DNA nanosensors detecting the mutation in the various cancers.

• Journal of the Optical Society of Korea
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• 제12권2호
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• pp.107-111
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• 2008

We demonstrated single-molecule fluorescence resonance energy transfer (FRET) from single donor-acceptor dye pair attached to a DNA with a setup based on a confocal microscope. Singlestrand DNAs were immobilized on a glass surface with suitable inter-dye distance. Energy transfer efficiency between the donor and the acceptor dyes attached to the DNA was measured with different lengths of DNA. Photobleaching of single dye molecule was observed and used as a sign of single-molecule detection. We could achieve high enough signal-to-noise ratio to detect the fluorescence from a single-molecule, which allows real-time observation of the distance change between single dye pairs in nanometer scale.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.142.1-142.1
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• 2003

Sensitive and safe method for visualization of DNA in agarose gels using visible dye is described. To improve the sensitivity, we studied a counterion-dye staining method using methyl orange as a counterion-dye which contributes to reduce excessive background staining by indoine blue. Dye concentrations, PH of staining solution, mixing molar ratio of two dyes, and staining times were optimized for the counterion-dye staining. By the staining with a mixed solution of 0.005% indoine blue and 0.00165% methyl orange in 10% ethanol 0.2M sodium acetate, 8 ng of the 3 kb DNA in an agarose gel was detected within 1hr. (omitted)

• Bulletin of the Korean Chemical Society
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• 제35권5호
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• pp.1455-1459
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• 2014

We report a sensitive and reliable FRET-based nanotechnology assay for efficient detection and quantification of bisulfite-unmodified or modified DNA. Bisulfite-untreated DNA or bisulfite-treated DNA is subjected to PCR amplification with biotin-conjugated primers so that the amounts of bisulfite-untreated and treated DNA can be differentiated. Streptavidin-coated quantum dots (QDs) are used to capture biotinylated PCR products intercalated with SYBR Green, enabling FRET measurement. Key features of our method include its low intrinsic background noise, high resolution, and high sensitivity, enabling detection of as little as 1.75 ng of bisulfite-untreated DNA in the presence of an approximately 1,000-fold excess of bisulfite-untreated DNA compared to bisulfate-treated DNA, with the use of PCR reduced (as low as 15 cycles). SYBR Green as an intercalating dye as well as a FRET acceptor allows for a single-step preparation without the need for primers or probes to be chemically conjugated to an organic fluorophore. Multiple acceptors per FRET donor significantly enhance the signal-to-noise ratio as well. In consideration of the high relevance of bisulfite treatment to DNA methylation quantitation, our system for FRET measurement between QDs and intercalating dyes can be generally utilized to analyze DNA methylation and to potentially benefit the scientific and clinical community.

• 융합정보논문지
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• 제8권5호
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• pp.45-50
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• 2018

형광 염료가 도핑 된 실리카 나노입자는 DNA 마이크로 에레이와 같은 바이오 라벨링 및 바이오 이미징에 활용되고 있으며, 높은 생체 적합성과 낮은 독성 및 높은 친수성의 특성을 가지고 있어 많은 주목을 받고 있는 기능성 나노소재이다. 본 논문에서는 형광 유기염료를 에탄올과 탈이온수에 각각 용해시킨 후 형광염료를 실리카 나노입자에 물리적으로 흡착시키는 방법과 화학적으로 도핑 시키는 방법으로 실리카 나노입자를 합성한 후 365 nm 파장의 자외선을 조사하여 형광특성을 분석하였다. 연구결과 형광 염료를 물리적으로 흡착시킨 실리카 나노입자보다 화학적으로 형광 염료를 도핑 시킨 실리카 나노입자의 형광특성이 우수하였으며, 도핑 된 형광 염료의 양이 많을수록 형광특성이 우수하였다. 형광 염료를 용해시키는 용매의 경우, 에탄올이 탈이온수와 비교하여 탁월한 형광 특성을 나타내었다. 또한 순수한 형광 염료와 형광 염료가 도핑된 실리카 나노입자의 광안정성을 조사한 결과, 형광 염료가 도핑 된 실리카 나노입자의 광안정성이 보다 우수한 것으로 나타났다. 이러한 연구결과를 바탕으로 형광염료가 최적으로 도핑 된 실리카 나노입자를 바이오 이미징 에이전트로 사용한다면 높은 광안정성과 형광특성으로 인하여 인체 내부의 생체 모니터링에 유용하게 활용될 것으로 전망된다.