• 제목/요약/키워드: Fluorometry

검색결과 44건 처리시간 0.023초

Rapid Detection of Ammonia-oxidizing Bacteria in Activated Sludge Based on 16S-rRNA Gene by Using PCR and Fluorometry

  • Hikuma, Motohiko;Nakajima, Masanori;Hirai, Toshiaki;Matsuoka, Hiroshi
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제7권5호
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    • pp.323-326
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    • 2002
  • To detect whole ammonia-oxidizing bacteria in the activated sludge, group-specific primers targeting the 16S-rRNA gene of ammonia-oxidizing bacteria were used. The electrophoresis pattern of the PCR products seemed to produce a single band of approximately 1.0 k bp for the bacteria in activated sludge and Nitrosomonas europaea. No band was observed for nitrite-oxidizer Nitrobacter winogradskyi and heterotrophs such as Pseudomonas putida. Then direct measurement of the PCR product was made by fluorometry using the reagent Hoechist 33258, so that the fluorescent intensity was in proportional to the cell number of the sample up to 240. Total time required for the test was about 4 h including DNA extraction. The DNA fragments produced were cloned and their sequences showed high similarity to those of Nitrosomonas spp. This study showed the feasibility to detect ammonia-oxidizing bacteria and to esti-mate their population rapidly for the control of the nitrogen elimination process.

Paper, Thin Layer 및 Column Chromatography에 의한 요중의 Estriol, Estrone, Estadiol-17 β의 분리 정량에 관하여 (Studies on Separation, Detection and Quantitation of Estriol, Estrone, Estradiol-17 β in Urine of Dairy Cows by Paper, Thin Layer and Column Chromatography)

  • 양용관;한수남;조종후
    • 대한수의학회지
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    • 제13권1호
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    • pp.23-30
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    • 1973
  • Thin layer, paper and column chromatography were compared for the separation, detection and quantitation of three kinds of estrogen in urine of dairy cows. While thin layer chromatography utilizing silica gel was better for the detection of estrogens, column chromatography using celite 545 was preferable. Spectrophotometry was compared with fluorometry for determination of estrone, estradiol-17 ${\beta}$ and estriol eluted by paper chromatography and column chromatography. Optical density of three standard estrogens showed almost same curve at maximum absorption wave length of 230 and $282m{\mu}$. However, the former showed a higher peak. In fluorometry, the fluorescence intensity of estrone and estradiol-17 ${\beta}$ were rather strong, when the estrogens were dissolved in sulfuric acid, and showed higher sensitivity than that of the spectrophotometry. However, in the case of estriol was exceptional.

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경상남도 함안군, 법수늪에서 엽록소 형광광도계(Phyto-PAM)에 의한 일차생산의 계절변동 (Seasonal Variations of Primary Productivity Analyzed by Phyto-PAM Chlorophyll Fluorometry in the Beopsu Marsh, Haman-gun, Gyeongsangnam-do)

  • 김미경
    • 환경생물
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    • 제26권2호
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    • pp.76-86
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    • 2008
  • 본 연구의 목적은 법수늪 유역 내에 유입되는 비점오염원과 늪지의 유입부의 점오염원 예측지역을 선정하여 수환경의 계절적 변화와 형광광도계(phyto-PAM)에 의한 1차 생산력을 분석하여 늪지 생태계의 관리방안을 위한 기초자료로 사용하고자 했다. 조사 시기는 2006년 2월, 4월, 8월과 11월에 각각 1회 정점 1$\sim$5의 수 표면에서 수심 1m사이의 수역에서 채수하고, 법수늪 유역 5개 지점, 늪 중앙을 1개 지점으로 하여 정점을 설정하였다. 특히, 생활하수와 오수정화시설에서 방류되는 하수로가 늪 안으로 유입되는 정점을 각각 2와 3으로 정하고, 인근의 청소년 수련원과 민가의 생활오수 등으로 유입되는 하수로 지점을 정점 4로 정하였는데 정점 3은 용존산소(1.39 mg L$^{-1}$)가 낮았고, 탁도, SS, T-N, T-P, BOD, COD, $Ca^{2+},\;Cl^-$가 장 높게 나타나 정점 6곳 중에 가장 오염원이 높은 곳으로 나타났다. BOD와 COD를 기준으로 했을 때 법수늪의 수질은 전반적으로 3급수 수준에 머물러 있다고 본다. 엽록소 ${\alpha}$의 농도는 4월에는 정점 2와 3에서 높게 나타났는데, 특히 정점 2에서는 급격히 증가하는 경향이 있었다(258.75 mg L$^{-1}$, 61.32 mg L$^{-1}$). 이는 4월에 정점 2에서 녹조류(Micratinium pusillum)가 급격히 증가했기 때문인 것으로 보인다. 식물플랑크톤의 2월에서 4월로 정점 2(180$\rightarrow$2,677 cells mL$^{-1}$)와 5(700$\rightarrow$6,590 cells mL$^{-1}$)에서 세포밀도가 크게 증가하였다. phyto-PAM에 의한 연중 식물플랑크톤의 분포도는 엽록소 a의 농도, 식물플랑크톤의 현존량과 높은 유의성이 높게 나타났다.

DYNAMICS OF $tRNA*{val}$ MEASURED WITH A LONG-LIFETIME METAL-LIGAND COMPLEX

  • Kang, Jung-Sook
    • Journal of Photoscience
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    • 제7권4호
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    • pp.155-159
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    • 2000
  • [Ru(bpy)$_2$(dppz)]$^2$$^{+}$ (bpy = 2,2'-bipyridine, dppz = dipyrido[3,2-a:2',3'-c]phenazine)(RuBD), a long-lifetime metal-ligand complex displays photophysical properties including long lifetime, polarized emission, and very little background fluorescence. To further show the usefulness of this luminophore(RuBD) for probing nucleic acid dynamics, its intensity and anisotropy decays when bound to tRN $A^{val}$ were examined using frequency-domain fluorometry with a blue light-emitting diode(LED)as the modulated light source. Unexpectedly much longer mean lifetime was obtained at 4$^{\circ}C$(<$\tau$>=178.3 ns) as compared to at $25^{\circ}C$(<$\tau$>=117.0 ns), suggesting more favorable conformation of tRN $A^{val}$ for RuBD when intercalated at 4$^{\circ}C$. The anisotropy decay data showed longer rotational correlation times at 4$^{\circ}C$(52.7 and 13.0 ns) than at $25^{\circ}C$ (32.9 and 10.3 ns). The presence of two rotational correlation times suggests that RuBD reveals both local and overall rotational motion of tRN $A^{val}$. Due to long lifetime of RuBD and small size of tRN $A^{val}$, very low steady-state anisotropy values were observed, 0.048 and 0.036 at 4 and $25^{\circ}C$, respectively. However, a clear difference in the modulated anisotropy values was seen between 4 and $25^{\circ}C$. These results indicate that RuBD can be useful for studying hydrodynamics of small nucleic acids such as tRN $A^{val}$.^{val}$.>.$.>.

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Fluorescence Resonance Energy Transfer in Calf Thymus DNA from a Long-Lifetime Metal-Ligand Complex to Nile Blue

  • Kang, Jung-Sook;Lakowicz, Josepb R.
    • BMB Reports
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    • 제34권6호
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    • pp.551-558
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    • 2001
  • We extended the measurable time scale of DNA dynamics to submicrosecond using a long-lifetime metal-ligand complex, $[Ru(phen)_2(dppz)]^{2+}$ (phen=1,10-phenanthroline, dppz=dipyrido[3,2-a:2',3'-c]phenazine) (RuPD), which displays a mean lifetime near 350 ns. We partially characterized the fluorescence resonance energy transfer (FRET) in calf thymus DNA from RuPD to nile blue (NB) using frequency-domain fluorometry with a high-intensity, blue light-emitting diode (LED) as the modulated light source. There was a significant overlap of the emission spectrum of the donor RuPD with the absorption spectrum of the acceptor NB. The F$\ddot{o}$rster distance ($R_0$) that was calculated from the spectral overlap was $33.4\;{\AA}$. We observed dramatic decreases in the steady-state fluorescence intensities of RuPD when the NB concentration was increased. The intensity decays of RuPD were matched the closest by a triple exponential decay. The mean decay time of RuPD in the absence of the acceptor NB was 350.7 ns. In a concentration-dependent manner, RuPD showed rapid intensity decay times upon adding NB. The mean decay time decreased to 184.6 ns at $100\;{\mu}M$ NB. The FRET efficiency values that are calculated from the mean decay times increased from 0.107 at $20\;{\mu}M$ NB to 0.474 at $100\;{\mu}M$ NB concentration. The use of FRET with a long-lifetime metal-ligand complex donor is expected to offer the opportunity to increase the information about the structure and dynamics of nucleic acids.

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해수 중 엽록소 a 측정방법에 대한 실험실 간 비교연구 (An Inter-Laboratory Comparison Study on Chlorophyll a Determination in Seawater)

  • 문초롱;강동진;박미옥;노재훈;유신재;문정언;신경훈;김윤숙;최중기;서영상
    • 한국해양학회지:바다
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    • 제19권1호
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    • pp.76-87
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    • 2014
  • 해양식물플랑크톤 생물량 및 일차생산력을 추정하기 위한 자료로서 이용되는 해수 중 엽록소 a의 농도는 고성능 액체크로마토그래피(High Performance Liquid Chromatography, HPLC), 형광법(Fluorometry), 분광광도법(Spectrophotometry)등의 분석방법으로 측정된다. 본 연구에서는 여러 실험실에서 이러한 방법들로 분석된 엽록소 a 자료를 통용하기 위한 기초자료로서 활용하고자 우리나라 주변 해역의 현장 시료를 이용한 상호비교실험을 실시하여 엽록소 a 분석 실험실 및 분석 방법간의 차이를 비교하였다. 상호비교실험은 엽록소 a 표준색소(R0) 및 동중국해(R1)와 동해해역(R2)에서 채수된 해수 현장 시료를 이용하여 총 3회 시행되었다. 참가실험실은 각각 HPLC(6개 실험실), 형광법(4개 실험실), 분광광도법(3개 실험실)으로 시료를 분석하였다. 표준색소와 현장시료의 측정결과에서 얻은 실험실 내 정밀도는 변동계수로 평가되었으며, 표준색소(R0)에서 9% 미만, 현장시료에서는 R1: 0.8~20%(평균 6.1%), R2: 4~21%(평균 13.2%)의 정밀도를 보였다. 전체 현장시료의 모집단에서 z-test를 이용하여 이상치를 제거한 측정결과들의 중앙값을 기준 값으로 평가한 HPLC, 형광법, 분광광도법 간의 차이는 20%이내였다. 이러한 차이는 현장시료의 균질성 및 실험실 내의 정밀도를 고려한 차이(R1: 8%, R2: 15%)와 유사한 값을 보였다. 비교결과로 미루어 볼 때, 측정값들간의 차이는 분석방법의 차이보다는 실험실 간 차이가 더 크다고 볼 수 있다. 따라서 각 실험실에서 생산된 자료는 분석 방법이 다르더라도 약 20% 이내에서는 동일한 결과를 생산한다고 볼 수 있으므로 타 실험실에서 서로 다른 방법으로 분석된 엽록소 a의 자료를 활용하는 경우에는 약 20%의 차이에 대한 고려가 필요하다.

Dynamics of RNA Bacteriophage MS2 Observed with a Long-Lifetime Metal-Ligand Complex

  • Kang, Jung Sook;Yoon, Ji Hye
    • Journal of Photoscience
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    • 제11권1호
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    • pp.35-40
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    • 2004
  • [Ru(2,2'-bipyridine)$_2$(4,4'-dicarboxy-2,2'-bipyridine)]$^{2+}$(RuBDc) is a very photostable probe that possesses favorable photophysical properties including long lifetime, high quantum yield, large Stokes' shift, and highly polarized emission. To evaluate the usefulness of this luminophore (RuBDc) for studying macromolecular dynamics, its intensity and anisotropy decays when conjugated to RNA bacteriophage MS2 were examined using frequency-domain fluorometry with a high-intensity, blue light-emitting diode (LED) as the modulated light source. The intensity decays were best fit by a sum of two exponentials, and the mean intensity decay time was 442.2 ns. The anisotropy decay data showed a single rotational correlation time (2334.9 ns), which is typical for a spherical molecule. The use of RuBDc enabled us to measure the rotational correlation time up to several microseconds. These results indicate that RuBDc can be useful for studying rotational diffusion of biological macromolecules.s.

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Ibuprofenlysine binding to human and bovine serum albumin using a fluorescence probe technique

  • Kim, Chong-Kook;Cha, Hyun-Sook;Kim, Yang-Bae;Yu, Byung-Sul
    • Archives of Pharmacal Research
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    • 제4권1호
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    • pp.19-24
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    • 1981
  • The possibility of using a fluorescence probe technique for the study of ibuprofenlysine binding to human and bovine serum albumin was investigated. 1-anilino-8-naphalenesulfonate was used as the probe. The number of binding sites of human and bovine serum albumins for ibuprofenlysine appears to be 4 and 2, respectively. By using this technique, the association constants were found to be $1.533{\times}10^{4}M^{-1}$ and $2.238{\times}10^{4}M^{-1}$, respectively.

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Surface Activity of Crude Ginseng Saponin

  • Kyu, Han-Suk;Kim, Nam-Hong
    • Archives of Pharmacal Research
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    • 제7권2호
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    • pp.109-113
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    • 1984
  • The critical micelle concentration (CMC) of crude ginseng saponin in water was determined by fluorometry and surface-tension measurement. These two methods gave the the CMC value, 0.015g/100ml AND 0.013G/100ml, respectively. The surface excess of the saponin and the area occupied by a saponin molecule at the monolayer adsorbed at air and waterinterface were calculated employing Gibbs adsorption equation. The presence of salt increased the surface activity of the saponin: it decreased the CMC, the surface tension at the CMC and the area occupied by a saponin molecule at the monolayer, which should be due to the salting-out effect of the salt.

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