• 제목/요약/키워드: 2D-gel

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칸나(Canna generalis)의 뿌리로부터 지질화합물의 분리.동정 (Isolation and Identification of Lipids from the Roots of Canna generalis)

  • 방면호;송명종;이대영;양혜정;한민우;백남인;이윤형
    • Applied Biological Chemistry
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    • 제49권4호
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    • pp.339-342
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    • 2006
  • 칸나의 뿌리를 80% MeOH용액으로 추출하고, 얻어진 추출물을 EtOAc, n-BuOH 및 물로 용매 분획 하였다. 이 중 EtOAc 분획과 n-BuOH 분획으로부터 silica gel과 octadecyl silica gel(ODS) column chromatography로 정제하여 4종의 화합물을 분리하였다. 각 화합물의 화학구조는 NHR, MS 및 IR 등의 스펙트럼 데이터를 해석하여, $\beta$-sitosterol(1), linoleic aicd methyl ester(2), monoglycosyldiglyceride(3), daucosterol(4)으로 동정하였다. 이 화합물들은 칸나에서 처음으로 분리되었다.

홍화(Carthamus tinctorius L.)로부터 활성물질의 분리 (Isolation of Biologically Active Compounds from the Flower Petals of Carthamus tinctorius L.)

  • 김융희;안은미;방면호;남지연;권병목;백남인
    • Applied Biological Chemistry
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    • 제41권2호
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    • pp.197-200
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    • 1998
  • 홍화 MeOH 추출물의 n-BuOH 분획으로부터 silica gel column chromatography를 반복하여 2종의 flavonoid 배당체를 분리, 정제하였다. 화합문의 화학구조를 HMBC를 포함한 NMR, MS, IR과 같은 기기분석 결과와 산가수분해반응을 이용하여 결정, 각각 $3-O-[{\beta}-D-glucopyranosyl(1{\rightarrow}2)\;{\beta}-D-glucopyranosyl]\;kaempferol$$3-O-[{\alpha}-L-rhamnopyranosyl(1{\rightarrow}6)\;{\beta}-D-glucopyranosyl]\;kaempferol$로 동정하였다. 각 화합물은 Grb2-Shc 결합저해활성$(IC_{50}:43,\;47\;{\mu}g/ml)$을 나타내었다.

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Neurospora crassa에서 알파아밀라제의 정제 및 유전자의 클로닝 (Purification and gene cloning of .alpha.-amylase of neurospora crassa)

  • 강일구;김미숙;양철학
    • 미생물학회지
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    • 제26권2호
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    • pp.73-81
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    • 1988
  • $\alpha$-Amylase (EC.3.2.1.1) of Neurospora crassa (ATCC9279) was cloned in E. coli HB101 using shotgun method, and the enzymes isolated from both N. crassa and E. coli were compared. Chromosomal DNA isolated from the spores of N. crassa was partially digested with PstI restriction endonuclease and rejoined to pBR322 which had been digested with the same enzyme. The resulting recombinant DNA were introduced into E. coli HB101 which had competancy by treating with $CaCl_{2}$. As the result, about 8000 colonies which showed tetracycline resistance were selected and two of the colonies which had 13.5Kb recombinant plasmid exhibit starch degrading activity on starch-containing plate when treated with D-cycloserine. $\alpha$-Amylases from both N.crassa and E. coli were isolated by using ammonium sulfate precipitation, DEAE-cellulose ion exchange column chromatography and Bio-Gel P150 gel foltration column. As the result, about 81.3 fold and 5.6 fold purifications in specific activities were obtained respectively, and specific activities of the gel filtrates were 6.1u/mg and 85u/mg respectively. The properties of both enzymes were compared and they showed quite the similar patterns in optimal temperature, optimal pH and had same molecular weight about 100,000 daltons on gel filtration method. Optimal temperatures for both enzymes were $70^{\circ}C$ and optimal pH were about 6 and 10.

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졸-겔법에 의한 Cugkadb 인산염계 수용성 유리의 제조 및 특성 (Preparation and Characterization of Water-Soluble Phosphate Glasses Containing Cu by Sol-Gel Method)

  • 오승환;최세영;김경남
    • 한국세라믹학회지
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    • 제35권4호
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    • pp.319-324
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    • 1998
  • Sol-gel derived phosphate water-sioluble glasses containing Cu were prepared. Powder-shape of glasses were added in D.I water used polyethylene bottle. After solution contained glass powder were submerged in water bath on 25$^{\circ}C$ their dissolution behavior/characteristics bactericidal effect and cytotoxicity test were evaluated. The maximum amount of Cu(35 mol%) via sol-gel method was more 5 mol% increased than that with melting process. The stage of total dissolution was more dominant than that of selective leaching dur-ing dissolution due to dissolved amount of glasses increased linearly with time. The ratio of Cu+ to {{{{ {Cu }^{2+ } }} was 3:7 so that the structure of glasses is more predominant 2-dimension chain structure than 3-dimenshion po-lymeric structue. The stage of total dissolution was more dominant than that of selective leaching during dissolution. Bactericidal effect against all bacteria showed that solutions which contained 40 ppm and 100 ppm of Cu killed 80 percentages of bacteria within 2 hours and 100 percentages of those within 12 hours. The results of cytotoxicity test for L929 cells showed no cytotoxicity were observed within 96 hours for dis-solved solution that contains 40 ppm and 100 ppm of Cu.

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Purification and Characterization of a Bacteriolytic Enzyme from Alkalophilic Bacillus sp.

  • Jung, Myeong-Ho;Kang, In-Soo;Bai, Dong-Hoon;Yu, Ju-Hyun
    • Journal of Microbiology and Biotechnology
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    • 제1권2호
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    • pp.102-110
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    • 1991
  • Alkalophilic Bacillus sp. YJ-451, which was isolated from soil at several area in Korea, produced a novel type of bacteriolytic enzyme (cell wall peptidoglycan hydrolase) extracellulary. The cell wall hydrolytic activity was identified as a clear zone on sodium dodecyl sulfate polyacrylamide gel electrophoresis containing 0.2% (w/v) cell wall of Bacillus sp. as substrate. This enzyme was successively purified 66 fold with 3.2% yield in culture broth by ammonium sulfate precipitation, CM-cellulose column chromatography, and gel filtration, followed by hydroxylapatite column chromatography. The molecular weight of the purified enzyme was estimated to be 27,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel filtration column chromatography. The optimum pH and temperature for the activity of the enzyme were pH 10.0 and $50^{\circ}C$, respectively. The enzyme was stable between pH 5.0 and 10.0 and up to $40^{\circ}C$. Among the microorganisms used in this experiment the enzyme was active against most of gram negative strains and the genus Bacillus such as B. megaterium, B. licheniformis, B. circulans, B. pumilus, B. macerans, B. polymyxa. The release of dinitrophenylglutamic acid but not reducing group from cell wall peptidoglycan digested by the enzyme suggested that the enzyme is a kind of peptidase which hydrolyzes the peptide bond at the amino group of D-glutamic acid in the peptidoglycan.

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Molecular Weight Determination of Polymers by Matrix Assisted Laser Desorption Ionization in Mass Spectrometry

  • Kim, Jin Sung;Yoo, Jong Shin
    • 분석과학
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    • 제8권4호
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    • pp.465-468
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    • 1995
  • Matrix assisted laser desorption ionization in mass spectrometry is a fast and accurate method to determine the molecular weight of natural and synthetic polymers. Unknown peptides such as elastase inhibitor and $\small{D}$-hydantoinase were analyzed using sinapinic acid as matrix and their molecular weights were compared with the results from protein sequencer and gel filtration chomatography, respectively. Synthetic polymers such as polyethyleneglycol, polypropyleneglycol, polydimethylsiloxane, and polystyrene were analyzed using matrices such as 2,5-dihydroxybenzoic acid, 4-hdroxyazobenzenecarboxylic acid, and 2-nitrophenyl octyl ether. Average molecular weights of polystyrene were compared with molecular weights by gel permeation chromatography.

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Evaluation of a Pretreatment Method for Two-Dimensional Gel Electrophoresis of Synovial Fluid Using Cartilage Oligomeric Matrix Protein as a Marker

  • Kong, Min-Kyung;Min, Byoung-Hyun;Lee, Pyung-Cheon
    • Journal of Microbiology and Biotechnology
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    • 제22권5호
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    • pp.654-658
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    • 2012
  • Osteoarthritis (OA) is the most common rheumatic pathology. One of the major objectives of OA research is the development of early diagnostic strategies such as those using proteomic technology. Synovial fluid (SF) in OA patients is a potential source of biomarkers for OA. The efficient and reliable preparation of SF proteomes is a critical step towards biomarker discovery. In this study, we have optimized a pretreatment method for twodimensional gel electrophoresis (2DE) separation of the SF proteome, by enriching low-abundance proteins and simultaneously removing hyaluronic acid, albumin, and IgG. SF samples pretreated using this optimized method were then evaluated by 1DE and 2DE separation followed by immunodetection of cartilage oligomeric matrix protein (COMP), a known OA biomarker, and by the identification of 3 proteins (apolipoprotein, haptoglobin precursor, and fibrinogen D fragment) that are related to joint diseases.

Expression Changes of Chironomus riparius Proteome reflecting Ecdysterone Inducible Genes and Mitogenome exposure to Bisphenol-A

  • Kwak Inn-Sil
    • 환경생물
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    • 제23권3호
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    • pp.275-280
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    • 2005
  • Development of the fourth-instar larvae of Chironomus riparius has a sensitive to ecdysteroid hormones. The 2D/E gel analysis for polypeptide expression reflecting early-ecdysterone inducible gene has conducted the emerged female from larval phase exposure to bisphenol A (BPA). In the 2D/E gel 1108 protein spots were identified. The visualized protein spots allowed extraction of 17 protein spots differed more than 3 fold in BPA treated animals, which was approximately $1.6\%$ of the total protein spots. However, polypeptide expression reflecting early-ecdysterone inducible gene didn't change after treatments. In addition, detection for the damages or changes in mitogenome level was observed. The conserved cytochrome oxidase I in DNA level affected exposure to BPA $(1{\mu}gL^{-1})$ in this preliminary study.

$PbTiO_3$-폴리머 O-3 복합 재료의 압전 특성에 관한 연구 (Piezoelectric Properties of O-3 Composite with $PbTiO_3$--Polymer)

  • 김일호;박인길;이성갑;이영희
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 1988년도 추계학술대회 논문집
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    • pp.50-53
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    • 1988
  • Lead titanate powders were prepared by mixed-oxide and sol-gel method. O-3 composites were prepared with these powders using Eccogel polymer. According to the volume percent of the PbTiO$_3$ powders, dielectric, piezoelectric properties and poling conditions were investigated. Relative dielectric constants were exponentially increased with volume percent of PbTiO$_3$. Piezoelectric strain coefficients (d$_{33}$ ) were increased with volume percent of PbTiO$_3$. Figure of merit (d$_{h}$.g$_{h}$) for the O-3 composites prepared by the mixed-oxide and sol-gel method were 2728(x10$^{-15}$ $m^2$/N) and 3100(x10$^{-15}$ $m^2$/N) at 70 volume percent of PbTiO$_3$, respectively.ely.

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Ferromagnetic Domain Behaviors in Mn doped ZnO Film

  • Soundararajan, Devaraj;Santoyo-Salazar, Jaime;Ko, Jang-Myoun;Kim, Ki-Hyeon
    • Journal of Magnetics
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    • 제16권3호
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    • pp.216-219
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    • 2011
  • Mn doped ZnO films were prepared on Si (100) substrates using sol-gel method. The prepared films were annealed at $550^{\circ}C$ for decomposition and oxidation of the precursors. XRD analysis revealed the presence of ZnMnO hexagonal wurtzite phase along with the presence of small quantity of $ZnMn_2O_3$ secondary phase and poor crystalline nature. The 2D, 3D views of magnetic domains and domain profiles were obtained using magnetic force microscopy at room temperature. Rectangular shaped domains with an average size of 4.16 nm were observed. Magnetic moment measurement as a function of magnetic field was measured using superconducting quantum interference device (SQUID) magnetometry at room temperature. The result showed the ferromagnetic hysteresis loop with a curie temperature higher than 300 K.