• 제목/요약/키워드: Fluorescence stability

검색결과 154건 처리시간 0.025초

Backbone Cyclization of Flavin Mononucleotide-Based Fluorescent Protein Increases Fluorescence and Stability

  • Tingting Lin;Yuanyuan Ge;Qing Gao;Di Zhang;Xiaofeng Chen;Yafang Hu;Jun Fan
    • Journal of Microbiology and Biotechnology
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    • 제33권12호
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    • pp.1681-1691
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    • 2023
  • Flavin mononucleotide-binding proteins or domains emit cyan-green fluorescence under aerobic and anaerobic conditions, but relatively low fluorescence and less thermostability limit their application as reporters. In this work, we incorporated the codon-optimized fluorescent protein from Chlamydomonas reinhardtii with two different linkers independently into the redox-responsive split intein construct, overexpressed the precursors in hyperoxic Escherichia coli SHuffle T7 strain, and cyclized the target proteins in vitro in the presence of the reducing agent. Compared with the purified linear protein, the cyclic protein with the short linker displayed enhanced fluorescence. In contrast, cyclized protein with incorporation of the long linker including the myc-tag and human rhinovirus 3C protease cleavable sequence emitted slightly increased fluorescence compared with the protein linearized with the protease cleavage. The cyclic protein with the short linker also exhibited increased thermal stability and exopeptidase resistance. Moreover, induction of the target proteins in an oxygen-deficient culture rendered fluorescent E. coli BL21 (DE3) cells brighter than those overexpressing the linear construct. Thus, the cyclic reporter can hopefully be used in certain thermophilic anaerobes.

수종 약물이 리포솜 지질막의 안정성에 미치는 영향 (Effects of Drugs on the Stability of Phospholipid Liposomal Membranes)

  • 김민;한석규;김종국
    • 약학회지
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    • 제38권6호
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    • pp.637-645
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    • 1994
  • The effect of various drugs on the stability of the liposomal membrane of phosphatidylcholine and cholesterol was studied, employing the fluorescence self-quenching method. Calcein was entrapped into the phospholipid small unilamellar vesicles and the leakage of the fluorescence probe was monitored on adding the drug to the system. The results of the experiments showed that phenothiazine derivatives, some potent local anesthetics and surface active agents were very effective in inducing the leakage of calcein from the liposome. The leakage-inducing activity of these drug substances has been ascribed to their surface activity and the perturbation of the liposomal membrane by these substances. On the other hand drug substance with low surface activity or without amphiphilic moieties did not show any effect or only small effect on the leakage of calcein from the liposomes. The effect of lipid concentration on the stability of the liposomes was also investigated to show that the higher concentrations of lipid more drug was required to induce the leakage. The effect of surface charges of vesicles was also studied, and the results showed that the charge on the liposomes enhanced the stability of the liposomes against the leakage-inducing activity of these drug substances.

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미세 성형 방법을 이용한 형광 표지된 이중 분획 입자의 제조 (Fabrication of Fluorescent Labeled Bi-compartmental Particles via the Micromolding Method)

  • 심규락;정성근;홍우경;강경구;이창수
    • Korean Chemical Engineering Research
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    • 제56권6호
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    • pp.826-831
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    • 2018
  • 본 연구는 다중 형광이 표지된 이중 분획 입자의 제조에 관한 것이다. 입자 내에서 형광 발현을 분획화하기 위하여, 형광의 여기 및 방출 스펙트럼의 중첩이 적은 두가지의 형광 염료를 선정한다. 또한, 형광 안정성을 확보하기 위하여 선정된 형광 염료는 입자를 구성하는 소재와 함께 가교될 수 있도록 분자 내에 아크릴레이트(acrylate) 작용기를 포함한다. 공초점 현미경 촬영을 통하여 선정된 형광 물질을 이용하여 제조된 입자에서 강한 형광 발현 및 형광의 분획화를 확인하였다. 더 나아가 4주 동안 형광 발현 및 세기를 측정하여 장기간의 형광 안정성을 검증하였다. 본 연구에서 제조된 다중 형광 표지된 이중 분획 입자는 다중 표적형 약물 전달 체계, 3차원 브라운 운동의 해석 연구, 3차원의 복잡한 자기 조립체 형상의 규명 연구 등에 널리 활용될 수 있으리라 기대한다.

자색고구마로부터 붉은 색소 농축물의 추출 및 그 색소 분말의 제조 특성 (Extraction of Natural Red Color Pigment Concentrate and Manufacturing Characteristics of Pigment Powder from Purple-Fleshed Sweet Potato)

  • 구본순;송대식
    • 한국식품영양학회지
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    • 제23권1호
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    • pp.23-29
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    • 2010
  • Pigment concentrates with violet-red color and sweet taste were obtained from purple-fleshed sweet potato(PFSP) using ethyl alcohol and water. Extract from general potato(GP) were used as a control. The relative stability of PFSP pigment concentrate(PFSPPC) in a storage test over 15 days was confirmed in the order of dark > fluorescence > sun-light irradiation. The relative stability of GP pigment concentrate(GPPC) in a storage test over 15 days was confirmed in the order of sun-light > fluorescence > dark storage. The RRP of PFSPPC was higher than that of GPPC, but the color strength of GPPC was 1/2 that of PFSPPC. Treatment of PFSPPC with aluminum potassium sulfate(0.2~0.3%, w/w) best improved its stability. The improved RRPs of PFSPPC were 45.16~47.31% in sun light irradiation, 55.91~60.22% in fluorescence irradiation, and 76.34~75.97% in dark storage conditions. In substituting aluminum potassium sulfate for chitosan, an amount of 0.2~0.3%(w/w) was suitable, giving similar results in improving pigment stability for all concentrates tested. Also, freeze-dried PFSPPC powder was manufactured as a substitute for dextrin, and also as a substitute for chitosan to the extent of 0.25%(w/w). The results of storage stabilite for freeze-dried PFSPPC and GPPC powder over 15 days, irradiation were, PRRs of 74.47~89.36% and 61.54~76.92%, respectively. The stability improving effect of freeze dried PFSPPC powder was confirmed by the results of storage experiments at various conditions. The use of freeze-dried PFSPPC powder was therefore confirmed to be an effective treatment for general foods.

제지용 형광증백제의 열 견뢰도에 대한 연구 (Study on the Thermal Fastness of Fluorescent Whitening Agents)

  • 김철환;이지영;김병호;최재성;임기백;김다미
    • 펄프종이기술
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    • 제44권1호
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    • pp.10-15
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    • 2012
  • Fluorescent whitening agents (FWAs) are used extensively in the paper industry to improve the optical properties. FWAs are neither photochemically nor thermally stable, and thereby the papers containing FWAs can be damaged by external heat. We observed that the CIE whiteness, ISO brightness and fluorescence index of the commercial papers decreased with the thermal treatment and thought that the thermal fastness of FWAs was the main factor affecting the thermal stability of the papers. Thus, the model papers treated with three types of FWAs were manufactured respectively and the thermal stability of FWAs was identified by determining ${\Delta}$ CIE whiteness and ${\Delta}$ fluorescence index.

속 이온과 CFP 상호작용: 흡수 형광 분광법에 의한 금속 이온과 CFP의 착물 형성 (Interaction of CFP with Metal ions: Complex Formation of CFP with Metal ion by Absorption and Fluorescence Spectrophotometery)

  • Siddiqi, K.S.;Mohd, Ayaz;Khan, Aftab Aslam Parwaz;Bano, Shaista
    • 대한화학회지
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    • 제53권2호
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    • pp.152-158
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    • 2009
  • 산성 용액에서 $Ca^{2+},\;Mg^{2+},\;Mn^{2+},\;Fe^{3+},\;Co^{2+},\;Ni^{2+},\;Cu^{2+}$$Zn^{2+}$와 Cefpodoxime proxetil (CFP)의 상 호작용을 분광학적으로 조사한 결과 1:1 착물이 형성됨을 알 수 있다. 순수한 약품의 흡수스펙트럼은 270과 345 nm에서 두 개의 현저한 봉오리를 보였다. 여러 pH에서 스펙트럼은 두 개의 isosbestic 점(305 과 330 nm)을 나타내었다. 이는 용액상에서 약품의 쯔비터 이온이 존재함을 의미한다. 다른 농도의 금 속이온에서 CFP의 형광방출 스펙트럼은 chelating enhancement fluorescence(CHEF)효과에 의해 형광강 도가 증가함을 알 수 있었다. 착체의 화학량론은 Job’'s 와 Benesi-Hildebrand 방법에 의해 결정되었다. 착 체의 안정도는 다음 순서와 같다. $Ca^{2+}\;<\;Mg^{2+}\;<\;Co^{2+}\;<\;Ni^{2+}\;<\;Zn^{2+}\;<\;Mn^{2+}\;<\;Cu^{2+}\;<\;Fe^{3+}$.

The Effect of the Oxygen Scavenging System on the pH of Buffered Sample Solutions: in the Context of Single-molecule Fluorescence Measurements

  • Kim, Sung-Eun;Lee, Il-Buem;Hong, Seok-Cheol
    • Bulletin of the Korean Chemical Society
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    • 제33권3호
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    • pp.958-962
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    • 2012
  • In single-molecule fluorescence experiment, the oxygen scavenging system is indispensable for avoiding photo-bleaching of fluorescent dyes. Here we report that the gloxy-based oxygen scavenging system commonly used in single molecule fluorescence experiments can disturb the solution pH considerably. To track in situ pH change, we utilized the pH-sensitive conformational transition of i-motif and examined the transition with ensemble and single-molecule FRET measurements. Based on our results, we also suggested several practical remedies for the stability of the solution pH.

Polyvinylalcohol에 고정화한 시금치 엽록체의 광합성특성에 대한 연구 (Photosynthetic Characteristics of Polyvinylalcohol-Immobilized Spinach Chloroplasts)

  • 박인호
    • Journal of Plant Biology
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    • 제34권3호
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    • pp.215-221
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    • 1991
  • Photoxynthetic properties of polyvinylalcohol (PVA)-immobilized chloroplast especially regarded to stability of photosynthetic electron transport and the fluorescence induction pattern were studied. When isolated spinach chloroplasts were immobilized with PVA, it showed good preservation of photosynthetic electron transport activity, especially PS II activity, during storage at -15$^{\circ}C$, 4$^{\circ}C$ and 2$0^{\circ}C$. And immobilized chloroplasts revealed similar thermostability of whole chain electron transport to free chloroplsts. And the absorption peak of red band of chloroplasts showed the blue-shift of 2-4 nm after immobilization. Fv/Fm ratio of chlorophyll fluorescence slightly decreased after immobilization. White light pulse after continuous light do not induced the additional fluorescence rise. This means chlorophyll fluorescence at room temperature reached to Fmax under continuous light in the immobilized chloroplasts. It seems that PVA may be a good candidate for immobilization matrix for the preservation of photosynthetic function of thylakoids and for the continuous use of chloroplast membranes of higher plants for solar energy storage and conversion.

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MTV가 연소실내 연료분포 및 화염 안정성에 미치는 영향 연구 (Effects of MTV on fuel distribution and flame stability in a SI engine)

  • 김기성
    • 한국분무공학회지
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    • 제2권4호
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    • pp.36-46
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    • 1997
  • The present study investigated the effects of MTV(Manifold Throttle Valve) on the fuel distribution and the flame stability in a SI engine at cold, idling condition. For the quantitative measurement of fuel distribution with PLIF method, compensating techniques of various factors such as laser beam nonuniformity, background image and local OTF nonuniformity were developed. As a result, it was found that MTV had a positive effect on the air-fuel mixing and flame stability.

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Acid Stability of Anti-Helicobacter pyroli IgY in in Aqueous Polyol Solution

  • Lee, Kyong-Ae;Chang, Sung-Keun;Lee, Yoon-Jin;Lee, Jong-Hwa;Koo, Nan-Sook
    • BMB Reports
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    • 제35권5호
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    • pp.488-493
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    • 2002
  • IgY was separated from a hen's egg yolk that was immunized with Helicobacter pyroli. The anti-H. pyroli IgY activity at acidic pH and the suppressive effect of polyol on acid-induced inactivation of IgY were investigated. Sorbitol and xylitol were used as polyols. IgY was quite stable at pH 5~7. Irreversible inactivation of IgY was observed at pH below 4, and proceeded rapidly at pH below 3. The acid stability of IgY was enhanced in the presence of 30% sorbitol or above. In a 50% aqueous sorbitol solution, an acid-induced inactivation was almost completely suppressed at pH 3. However, the improvement of IgY activity was not observed in the aqueous xylitol solution. IgY showed almost the same activity as native IgY when sucrose was substituted for sorbitol. On the other hand, the xylitol replacement with sucrose did not enhance the acid stability of IgY. The acid-induced inactivation of IgY was related to tryptophyl fluorescence. Fluorescence emission spectra suggested that structural changes near the tryptophan residues may occur under acidic conditions. An increase in sorbitol concentration induced a blue shift. The fluorescence emission of IgY in a 50% sorbitol solution had a peak at 330 nm, which was the same emission peak that was exhibited by native IgY. Sorbitol could, therefore, be used as a good stabilizer of IgY under acidic conditions.