• Title/Summary/Keyword: Agarose

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Directed Colony Hybridization Using Agarose Gel

  • Park, Jong-Chun;Chun, Soon-Bai
    • Journal of Microbiology and Biotechnology
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    • v.4 no.3
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    • pp.235-236
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    • 1994
  • Direct colony hybridization on agarose gel plate was established for the identification of recombinant plasmids. The hybridization of the probe to nucleic acids on dried gel without transferring to solid supports was more effective and simpler than hybridization of such probes to materials immobilized on filters such as nitrocellulose or nylon. D-cycloserine in overlaying agamse was essential for releasing the nucleic acids from colony.

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Nucleotide Binding Component of the Respiratory Burst Oxidase of Human Neutrophils

  • Park, Jeen-Woo;Ahn, Soo-Mi
    • BMB Reports
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    • v.28 no.3
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    • pp.275-280
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    • 1995
  • The respiratory burst oxidase of neutrophils is a multicomponent enzyme, domant in resting cells, that catalyzes the reduction of oxygen to $O_{2}^{-}$ at the expense of NADPH. In the resting neutrophil, some of the components of the oxidase, including proteins p47 and p67, are in the cytosol, while the rest are in the plasma membrane. Recent evidence has suggested that at least some of the cytosolic oxidase components exist as a complex. The cytosolic complex with a molecular weight of ~240 kDa was found to bind to blue-agarose and 2',5'-ADP-agarose, which recognize nucleotide requiring enzymes. In order to identify the nucleotide binding component of the cytosolic complex we purified recombinant p47 and p67 fusion proteins using the pGEX system. Pure recombinant p47 was retained completely on 2',5'-ADP-agarose, whereas pure recombinant p67 did not bind to these affinity beads. On the basis of these results, we infer that p47 may contain the nucleotide binding site.

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PHOTOEVOLUTION OF HYDROGEN FROM AQUEOUS ALCOHOL WITH AGAROSE GEL SYSTEM CONTAINING CdS or $TiO_2$PARTICLES

  • Park,Yong-Tae;Lee, Sang-Gyun;Cheong, Jong-Jae;Lee, Jai-Young
    • Journal of Photoscience
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    • v.5 no.1
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    • pp.27-31
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    • 1998
  • An artificial photolysis system of water or aqueous alcohol is studied. Upon irradiation of agarose gel system containing colloidal CdS or TiO$_2$ particles with Rh catalyst, hydrogen and oxygen are produced from water. When an alcohol is used as an electron donor in the agarose system, hydrogen and oxidized form of alcohol, the corresponding an aldehyde, is produced. The photolysis mechanism of water or alcohol in the agarose system is proposed.

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Development of Cosmetic Emulsion Using Blueberry Fruit Extract and Agarose from Gracilaria verrucosa (꼬시래기 유래 아가로즈와 블루베리 열매 추출물을 이용한 화장용 에멀젼 개발)

  • Choi, Moon-Hee;Kim, Yong-Woon;Kim, Mi-Sook;Shin, Hyun-Jae
    • KSBB Journal
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    • v.31 no.4
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    • pp.256-262
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    • 2016
  • The need for natural cosmetic ingredients has been increasing over the world nowadays. Agarose, a natural polymer from red seaweeds, has high hydrophilic character and a function of scaffolder. As skin moisturizer, agarose is adequate for percutaneous absorption. While, blueberry fruits extract possesses rich procyanidins and anthocyanins which show health benefits, anti-oxidant effect, anti-aging and anti-melanogenesis. Stability, sensory preference, skin trouble of the emulsion formula are important for cosmetic product development. In this study, we manufactured an emulsion formula for skin moisturizers using the two ingredients and tested emulsion stability and skin trouble. Total phenolic contents of the blueberry fruits extract were evaluated as well as tyrosinase inhibitory and collagenase inhibitory activities. $IC_{50}$ values of blueberry fruits extract for anti-tyrosinase and anti-collagenase activities were 168 and $112{\mu}g/mL$, respectively using gallic acid as a control ($64.8{\mu}g/mL$). The stability (pH and viscosity) of the formula containing 2% blueberry fruits extracts and 0.1% agarose was measured at five different temperatures (room temp., $25^{\circ}C$, $55^{\circ}C$, $45^{\circ}C$, $55^{\circ}C$) under the sun light at 2 day intervals for 12 days. There has been little pH change at the different temperatures. According to the sensory evaluation, there was no significant flavor, discoloration and physical changes of the formula at $25-65^{\circ}C$. These results suggest that emulsion formula containing blueberry extract and agarose could be used as a candidate for lotion and essence products.

Microcontact Printing of Bacteria Using Hybrid Agarose Gel Stamp (혼성 아가로즈젤 스탬프를 이용한 박테리아 마이크로 컨택트 프린팅)

  • Shim, Hyun-Woo;Lee, Ji-Hye;Lee, Chang-Soo
    • KSBB Journal
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    • v.21 no.4
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    • pp.273-278
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    • 2006
  • The noble method of hybrid agarose gel microstamp fabricated by replica molding against PDMS master to make bacteria patterns on agar surface was presented. After the fabricated hybrid agarose gel microstamp was inked with E. coli, we could obtain 2 dimensional bacterial arrays with $50{\mu}m$ circular spots. And the various shaped patterns based on experimental design were easily generated. The analysis of mean fluorescent signal was showed that bacterial pattern have high contrast between spots and background and homogeneity of pattern. Our proposed method solved the problem of wetting and handling with small soft agarose gel microstamp when bacteria were used for ink. The agarose gel stamp provides appropriate environment to inked bacteria, which is essential technology for cell patterning with high retaining viability during the patterning process. This method is reproducible, convenient, rapid, and could be applied to screening system, study of cell-surface interaction, and microbial ecology.

Immobilization of Endo- and Exoinulinase on Vinylsulfone Activated Agarose (Vinylsulfone Activated Agarose 에 Endo- 및 Exoinulinase의 고정화)

  • 한상배;송근섭;정용섭;손희숙;우순자;엄태봉
    • Microbiology and Biotechnology Letters
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    • v.20 no.1
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    • pp.20-24
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    • 1992
  • In order to reuse inulinase effectively, a method for immobilizing both endo- and exoinulinase to vinylsulfone activated agarose via covalent bond was investigated. The immobilized enzyme preparation had, respectively, 400 U for exoinulinase activity and 80 U for endoinu- Iinase activity per gram gel. A thermal stability by immobilization had increased in the case of exoinulinase. Optimum pHs for two immobilized enzymes were 4.4 to 5.0. Synergistic effect which depends on mixed ratio of two immobilized enzymes was the best when the mixed ratio of endo/exo lay between 0.1 and 0.5, and its activity of the mixed enzyme increased 1.7 times as compared to that of each immobilized enzyme. Inulinase activities of both of the immobilized enzymes did not change during 20 times experimental runs in a batch reactor.

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Production of Reuterin by Immobilized Lactobacillus reuteri (Lactobacillus reuteri의 고정화 세포를 이용한 루테린 생산)

  • Yum, Eun-Mi;Noh, Bong-Soo;Ji, Geun-Eog
    • Korean Journal of Food Science and Technology
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    • v.37 no.2
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    • pp.318-320
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    • 2005
  • Lactobacillus reuteri residing in human and animal intestines converts glycerol into reuterin (antimicrobial substance) in anaerobic condition. Attempt was made to increase production efficiency of L. reuteri by employing immobilized cells. L. reuteri was immobilized in agarose beads, which were then reacted with 250 mM glycerol solution. Batch-type production of reuterin with immobilized cells (0.5% agarose beads) lasted for about 36 h, although reuterin production decreased with passage of time. In continuous-type production, period of reuterin production with immobilized cells was extended about twofold and production ratio increased 1.5-fold (502 mM) compared with suspended cells (315 mM). Maximum concentration of reuterin reached 47 mM at 80 min after reaction with glycerol solution. Results of this study indicate that immobilization of Lactobacillus reuteri in agarose beads increased reuterin production.

Immobilization of Recombinant Bacterial Biosensors: a Simple Approach for the On-Site Detection of Phenolic Compounds (재조합 박테리아 바이오센서의 고정화: 페놀계 화합물의 현장 검출을 위한 간단한 접근 방법)

  • Shin, Hae-Ja
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1323-1328
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    • 2011
  • We herein report the development of an agarose-gel-immobilized recombinant bacterial biosensor simple system for the field monitoring of phenolic compounds. Escherichia coli cells harboring the pLZCapR plasmid, which was previously designed to express the ${\beta}$-galactosidase reporter gene in the presence of phenolic compounds, were co-immobilized with a substrate [chlorophenol red ${\beta}$-galactopyranoside (CPRG) in agarose gel, and dispensed to the wells of a 96-well plate. Field samples were added to the wells and color development was monitored. In the presence of 5 ${\mu}M$ to 10 mM of phenol, the biosensor developed a red (representing hydrolysis of CPRG) color. Other phenolic compounds were also detected by this immobilized system, with the pattern resembling that previously reported for the corresponding non-immobilized biosensor. The immobilized cells showed optimum activity when the gel was simultaneously supplemented with 6% dimethyl formamide (DMF), 0.1% SDS and 10 mM $CaCl_2$. The immobilized biosensor described herein does not require the addition of a substrate or the use of unwieldy instruments or sample pretreatments that could complicate field studies.

Separation of Single-Wall Carbon Nanotubes by Agarose Gel (아가로스 겔을 이용한 단일벽 탄소나노튜브 분리)

  • Yu, Lan;Lim, Yun-Soo;Han, Jong-Hun
    • Applied Chemistry for Engineering
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    • v.22 no.3
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    • pp.272-276
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    • 2011
  • The separation of metallic and semiconducting single-wall carbon nanobubes (SWCNTs) by agarose gel method was carried out in this study. The effect of concentration of agarose, SDS (sodium dodecyl sulfate), and pH in the solution on separation behavior was investigated. With increasing the concentration of agarose in the solution, it showed that the ratio of metallic SWCNTs, which was analyzed from UV-vis-NIR spectroscopy, was increased in the solution phase, while the overall concentration of SWCNTs was decreased. With increasing the concentration of SDS, we could observe that the ratio of metallic SWCNTs was increased due to more affinity between SDS molecules and metallic SWCNT. The highest metallic SWCNTs ratio was reached up to 58.4% when the pH of solution was 8.2.

Evaluation of polyglycolic acid as an animal-free biomaterial for three-dimensional culture of human endometrial cells

  • Sadegh Amiri;Zohreh Bagher;Azadeh Akbari Sene;Reza Aflatoonian;Mehdi Mehdizadeh;Peiman Broki Milan;Leila Ghazizadeh;Mahnaz Ashrafi;FatemehSadat Amjadi
    • Clinical and Experimental Reproductive Medicine
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    • v.49 no.4
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    • pp.259-269
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    • 2022
  • Objective: Animal-free scaffolds have emerged as a potential foundation for consistent, chemically defined, and low-cost materials. Because of its good potential for high biocompatibility with reproductive tissues and well-characterized scaffold design, we investigated whether polyglycolic acid (PGA) could be used as an animal-free scaffold instead of natural fibrin-agarose, which has been used successfully for three-dimensional human endometrial cell culture. Methods: Isolated primary endometrial cells was cultured on fibrin-agarose and PGA polymers and evaluated various design parameters, such as scaffold porosity and mean fiber diameter. Cytotoxicity, scanning electron microscopy (SEM), and immunostaining experiments were conducted to examine cell activity on fabricated scaffolds. Results: The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay and SEM results showed that endometrial cells grew and proliferated on both scaffolds. Immunostaining showed cytokeratin and vimentin expression in seeded cells after 7 days of culture. On both scaffolds, an epithelial arrangement of cultured cells was found on the top layer and stromal arrangement matrix on the bottom layer of the scaffolds. Therefore, fibrin-agarose and PGA scaffolds successfully mimicked the human endometrium in a way suitable for in vitro analysis. Conclusion: Both fibrin-agarose and PGA scaffolds could be used to simulate endometrial structures. However, because of environmental and ethical concerns and the low cost of synthetic polymers, we recommend using PGA as a synthetic polymer for scaffolding in research instead of natural biomaterials.