• 제목/요약/키워드: Fluorescence quenching

검색결과 239건 처리시간 0.024초

산화 그래핀 플랫폼을 이용한 DNA 중합효소의 실시간 형광에세이 (Real-time Fluorescence Assay of DNA Polymerase Using a Graphene Oxide Platform)

  • 강종백
    • 한국미생물·생명공학회지
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    • 제41권4호
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    • pp.456-461
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    • 2013
  • 단일가닥 DNA와 이중가닥 DNA의 흡착 율의 차이를 이용하여, 본 연구는 산화 그래핀에 흡착된 단일 가닥 DNA을 사용하여 Klenow fragment의 효소 활성을 검출하기 위하여 실시간 형광에세이 방법을 사용했다. 실험 결과에 의하면, 산화그래핀에 흡착된 형광표지 ssDNA는 형광이 ��칭(quenching) 되지만, cDNA 첨가에 의해서 흡착된 단일가닥 DNA가 유리되었다. Klenow fragment의 활성을 측정하기 위해서 형광표지 틀(template) DNA, 산화그래핀과 시발체(primer)가 존재할 때, 고분자 반응이 진행됨에 따라 ��칭된 형광세기가 증가하였다. 그리고 겔 전기영동 실험은 산화 그래핀에서 DNA 합성과 hybridization 반응을 확인하였다.

Interaction of Bilobalide and Ginkgolides B with Bovine Serum Albumin: A Fluorescence Quenching Study

  • Chen, Yan;Wang, Ruijun;Wang, Shusheng;Yang, Yi;Li, Shaofei;Kai, Guiqing
    • Bulletin of the Korean Chemical Society
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    • 제32권9호
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    • pp.3261-3266
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    • 2011
  • The interaction of bilobalide (BB) and ginkgolides B (GB) with bovine serum albumin (BSA) was investigated by fluorescent technique and UV/vis absorption spectroscopy. The results showed that BB and GB could intensively quench the fluorescence of BSA through a static quenching procedure. The binding constants (Ka) and the average binding distance between the donor (BSA) and the acceptor (ginkgolides) were obtained ($r_{BB}$ = 5.33 nm and $r_{GB}$ = 4.20 nm) by the theory of non-radiation energy transfer, and then the thermodynamic parameters such as ${\Delta}S^0$ (0.17-0.32 kJ/mol), ${\Delta}G^0$ (-20.76 ~ -17.79 kJ/mol) and ${\Delta}H^0$ (32.47-76.52 kJ/mol) could be calculated, respectively. All these results revealed that the interaction of BB and GB with BSA were driven mainly by hydrophobie force. The synchronous fluorescence spectroscopy was applied to examine the effect of two ginkgolides on the configuration of BSA. The configuration alteration of BSA could be induced by the hydrophobicitv environment of tyrosine with the increase of the drug concentration.

Effects of Parathyroid Hormone on the Fluidity of the Plasma Membrane Vesicles of Cultured Osteoblasts

  • Kang, Jung-Sook
    • Journal of Photoscience
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    • 제8권3_4호
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    • pp.87-92
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    • 2001
  • Intramolecular excimer formation of 1,3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to investigate the effects of parathyroid hormone (PTH) on the bulk bilayer fluidity of the plasma membrane vesicles isolated from cultured osteoblasts (OB-PMV). In a dose-dependent manner, rat PTH-(1-34) [rPTH-(1-34)] increased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py and decreased the anisotropy (r) of DPH in OB-PMV. This indicates that PTH increased both the lateral and rotational diffusion of the probes in OB-PMY. Selective quenching of DPH fluorescence by trinitrophenyl groups was utilized to examine the transbilayer fluidity asymmetry of OB-PMV. The anisotropy, limiting anisotropy, and order parameter of DPH in the inner monolayer were 0.024, 0.032, and 0.062 greater than calculated for the outer monolayer of OB-PMY. Selective quenching of DPH fluorescence by trinitrophenyl groups was also utilized to examine the transbilayer effects of PTH on the fluidity of OB-PMV. rPTH-(1-34) had a greater fluidizing effect on the outer monolayer as compared to the inner monolayer of OB-PMV. Thus, it has been proven that PTH exhibits a selective rather than nonselective fluidizing effect within transbilayer domains of OB-PMV.

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PHOTOPHYSICAL PROPERTIES OF FLUORENONES WITH CHIRAL SUBSTITUENTS AND THEIR ASYMMETRIC RECOGNITION THROUGH INTERMOLECULAR HYDROGEN BONDING INTERACTIONS IN THE EXCITED STATES

  • Aikawa, Yoshihide;Shimada, Tetsuya;Tachibana, Hiroshi;Inoue, Haruo
    • Journal of Photoscience
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    • 제6권4호
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    • pp.165-170
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    • 1999
  • Asymmetric recognition of chiral alcohol by fluorenone derivatives with chiral substituents through intermolecular hydrogen bonding interaction in the singlet excited state was attempted. 1-((1S, 2R, 5S)-(+)-Menthyloxycarbonyl)aminofluoren-9-one (1-MAF) and 1-((1S, 2R, 5S)-(+)-menthyloxycarbonyl)oxyfluoren-9-one (1-MOF) were synthesized and their photophysical behaviors were characterized by the measurement of absorption and fluorescence spectra, as well as the quantum yield and the lifetime of fluorescence. The excited singlet states of 1-MAF and 1-MOF were revealed to have characteristics similar to those of fluorenone, though the intramolecular CT nature was fairly suppressed as compared with 3- and 4-substituted aminofluorenones. Fluorescences of 1-MAF and 1-MOF in acetonitrile were quenched by the addition of alcohols. Differences in fluorescence quenching efficiency were hardly observe for rather small chiral alcohols such as (R)-(-)- or (S)-(+)-2-butanol, while bulky alcohols such as menthol and isopinocampheol showed chiral recognition effects in their fluorescence quenching of 1-MAF in either acetonitrile or butyronitrile.

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Investigation of effects of newly synthesized Pt(II) complex against human serum albumin and leukemia cell line of K562

  • Divsalar, Adeleh;Saboury, Ali A.;Ahadi, Leila;Zemanatiyar, Elham;Mansouri-Torshizi, Hassan
    • BMB Reports
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    • 제43권11호
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    • pp.766-771
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    • 2010
  • The biological evaluation of a new synthesized Pt(II)-complex, 2,2'-bipyridin Butylglycinato Pt(II) nitrate, an anti-tumor component, was studied at different temperatures by fluorescence and far UV circular dichroism (CD) spectroscopic methods. Human serum albumin (HSA) and human tumor cell line K562 were as targets. The Pt(II)-complex has a strong ability to quench the intrinsic fluorescence of HSA. Binding and thermodynamic parameters of the interaction were calculated by fluorescence quenching method. Far-UV-CD results showed that Pt(II)-complex induced increasing in content of $\alpha$ helical structure of the protein and stabilized it. The 50% cytotoxic concentration ($Cc_{50}$) of complex was determined using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay at different incubation times. Also, fluorescence staining with DAPI (4,6-diamidino-2-phenylindole) revealed some typical nuclear changes, which are characteristic of apoptosis. Above results suggest that Pt (II) complex is a promising anti-proliferative agent and should execute its biological effects by inducing apoptosis.

Various Sensor Applications Based on Conjugated Polymers

  • Lee, Chang-Lyoul
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2014년도 제46회 동계 정기학술대회 초록집
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    • pp.103.1-103.1
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    • 2014
  • Due to their excellent optical and electrochemical properties, conjugated polymers have attracted much attention over the last two decades and employed to opto-electrical devices. In particular, conjugated polymers possess many attractive features that make them suitable for a variety of sensing task. For example, their delocalized electronic structures can be strongly modified by varying the surrounding environment, which significantly affected molecular energy level. In other word, conjugated polymers can detect and transduce the environmental information into a fluorescence signal. Conjugated polymers also display amplified quenching compared to small molecule counterparts. This amplified fluorescence quenching is attributed to the delocalization and migration of the excitons along the conjugated polymer backbones. Long backbones of conjugated polymer provide the transporting path for electron as a conduit, allowing that excitons migrate rapidly into quencher site along the backbone. This is often referred to as the molecular wire effect or antenna effect. Moreover, structures of conjugated polymers can be easily tailored to adjust solubility, absorption/emission properties, and regulation of electron/energy transfer. Based on this versatility, conjugated polymers have been utilized to many novel sensory platforms as a promising material. In this tutorial, I will highlight a variety of fluorescence sensors base on conjugated polymer and explain their sensory mechanism together with selected examples from reference literatures.

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보리 엽록체의 광계 II에서 수은, 구리 및 아연의 저해효과 비교 (Comparison of Toxic Effects of Mercury, Copper and Zinc on Photosystem II of Barley Cholroplasts)

  • 전현식
    • Journal of Plant Biology
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    • 제36권3호
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    • pp.195-201
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    • 1993
  • The room temperature fluorescence induction of chloroplasts was utilized as a probe to locate the site of inhibition by mercury, copper and zinc on PS II by mercury. Inhibitory effect of Hg2+ on electron transport activity was notable as compared with Cu2+ and Zn2+. At concentrations of HgCl2 over 50 $\mu$M, activities of PS II and whole-chain electron transport decreased more than 70%, while that of PS I decreased about 10~30%. This suggests that PS II is more susceptible to Hg2+ than PS I is. In the presence of diphenylcarbazide (DPC), 50 $\mu$M HgCl2 inhibited the reduction of dichlorophenolindophenol (DCPIP) about 50%. Addition of heavy metals induced marked decrease in maximal variable fluorescence/initial fluorescence [(Fv)m/Fo], but no changes in Fo. With various concentrations of heavy metals, changes of chlorophyll a fluorescence emitted by PS II showed gradual decrease in photochemical quenching (qQ), which indicates an increase in reduced state of electron acceptor, QA. Especially, the addition of HgCl2 caused a notable decrease of qQ. In the presence of 50 $\mu$M CuCl2, energy-depended quenching (qE) was completely reduced, whereas in the presence of 50 $\mu$M CuCl2 and ZnCl2 it was still remained. The above results are discussed on the effects of mercury in relation to water-splitting system and plastoquinone (PQ) shuttle system.

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Photocycloaddition Reaction of 1,2-Bispyrazylethylene to Tetracyanoethylene

  • Shim, Sang-Chul;Shim, Hyun-Kwan
    • Bulletin of the Korean Chemical Society
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    • 제1권4호
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    • pp.123-126
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    • 1980
  • Benzene solution of trans-1,2-bispyrazylethylene and electron deficient olefin, tetracyanoethylene, as a ${\pi}$-acceptor gave 1,2-bispyrazyl-3,3,4,4-tetracyanocyclobutane, a 2${\pi}$ + 2${\pi}$ cycloaddition product, on irradiation with 350 nm UV light. Fluorescence studies revealed the reaction to proceed through a singlet exciplex. The fluorescence of trans-1,2-bispyrazylethylene was quenched very efficiently by tetracyanoethylene with the quenching constant of 1.6 ${\times}$ 10$^{10}$M$^{-1}$s$^{-1}$ while electron rich olefin, tetramethylethylene, did not quench the fluorescence of bispyrazylethylene.

고추의 엽록소 형광 이미지 분석법에 의한 한발스트레스 지표화 가능성 (Possibility of Drought stress Indexing by Chlorophyll Fluorescence Imaging Technique in Red Pepper (Capsicum annuum L.))

  • 유성녕;엄기철;박소현;김태완
    • 한국토양비료학회지
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    • 제45권5호
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    • pp.676-682
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    • 2012
  • 본 연구에서는 생육초기 고추의 한발 스트레스 조건에서 작물 생육에 미치는 영향을 구명하기 위해 엽록소 이미지형 광분석을 이용하여 엽록소 형광 매개변수를 비교분석 하였다. 비록 통계적 유의성이 인정되지는 않았지만 전체 잎의 이미지에 나타나는 형광매개변수들의 평균값은 감소하는 경향이었다. 가변형광값에대한 최대형광값의 비는 변동이 없는 반면, 광계II의 정류상태에서의 양자수득율 및 형광감쇄율은 다소 감소하였다. 이러한 현상은 퀴논 A 단백질로부터 퀴논 B 단백질로의 전자전달의 감소를 의미하였다. 잎의 중앙부위와 엽병부근에서의 비광학적 소광 및 가변형광의 비광학적 소광 상수는 한발스트레스 4일 후 유의성있게 증가하였다. 즉 생리적 스트레스 매개변수로 활용이 가능하였다. 광계II에서의 정류상태 양자수득율의 감소는 고추잎의 모든 부위에서 공통적으로 유의성있게 측정되었다. 결론적으로 ${\Phi}PSII$ ($QY_{_-LSS}$), $NPQ_{_-LSS}$, qN 등의 엽록소형광 매개변수들은 고추의 한발스트레스를 판단할 수 있는 생리적 지표로 활용 가능한 것으로 유의성이 인정되었다.