• Title/Summary/Keyword: Extraction buffer

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Biochemical Properties and Localization of the β-Expansin OsEXPB3 in Rice (Oryza sativa L.)

  • Lee, Yi;Choi, Dongsu
    • Molecules and Cells
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    • v.20 no.1
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    • pp.119-126
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    • 2005
  • ${\alpha}$-Expansins are bound to the cell wall of plants and can be solubilized with an extraction buffer containing 1 M NaCl. Localization of ${\alpha}$-expansins in the cell wall was confirmed by immunogold labeling and electron microscopy. The subcellular localization of vegetative ${\beta}$-expansins has not yet been studied. Using antibodies specific for OsEXPB3, a vegetative ${\beta}$-expansin of rice (Oryza sativa L.), we found that OsEXPB3 is tightly bound to the cell wall and, unlike ${\alpha}$-expansins, cannot be solubilized with extraction buffer containing 1 M NaCl. OsEXPB3 protein could only be extracted with buffer containing SDS. The subcellular localization of the OsEXPB3 protein was confirmed by immunogold labeling and electron microscopy. Gold particles were mainly distributed over the primary cell walls. Immunohistochemistry showed that OsEXPB3 is present in all regions of the coleoptile and root tissues tested.

Separation of Lipase Using Reverse Micelles in Spray Column (Spray Column에서 역미셀을 이용한 Lipase의 분리)

  • 한동훈;홍원희
    • KSBB Journal
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    • v.8 no.1
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    • pp.83-88
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    • 1993
  • Lipase was separated using reverse mlcelles in a spray column. The 50 mM AOT-Isooctane solution was used as reverse micellar solution for the extraction of lipase (crude containing 25% Protein). Ionic strength was controlled by KCl(0.1M KCl for extraction, 0.5M KCl for back exlractlon). Acetate buffer and phosphate buffer were used for control of pH. The efficiencies of extraction and stripping were 30% and 50%. An increase of circulation did not change the efficiency of extraction in forward extraction. The optimum flow rate was around 0.10ml/sec.

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The Effects of pH Change in Extraction Solution on the Heavy Metals Extraction from Soil and Controversial Points for Partial Extraction in Korean Standard Method (용출액의 pH 변화가 토양내 중금속 용출에 미치는 영향과 그에 따른 국내 토양 오염 공정시험방법의 문제점)

  • 오창환;유연희;이평구;이영엽
    • Economic and Environmental Geology
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    • v.36 no.3
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    • pp.159-170
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    • 2003
  • Heavy metals are extracted from Chonju stream sediment, roadside soils and sediments along Honam expressway, soils and tailings from mining area using three different methods (partial extraction in Standard Method, partial extraction method with maintaining 0.1 N of extraction solution and Sequential Extraction Method). In samples having buffer capacity against acid, pH 1 (0.1 N HCl) of extraction solution can not be maintained and pH of extraction solution increases up to 8.0 when partial extraction in Standard Method is used. The averages and ranges of HPE(heavy metals extracted using partial extraction in Standard Method)/HPEM(heavy metals extracted using partial extraction method with maintaining 0.1 N of extraction solution) values are 0.479 and 0.145~0.929 for Cd, 0.534 and 0.078~0.928 for Zn, 0.432 and 0.041~0.992 for Mn, 0.359 and 0.011~0.874 for Cu, 0.150 and 0.018~0.530 for Cr, 0.219 and 0.003~0.853 for Pb, and 0.088 and 1.73${\times}$10$^{-5}$~0.303 for Fe. These data indicate that the difference between HPE and HPEM is large in the order of Fe, Cr, Pb, Cu, Mn, Cd and Zn. The amounts of heavy metals extracted decreases in the follow order; Sum III(sum of fraction I, II, III in sequential extraction)>HPEM>Sum III (sum of fraction I and II)>HPE for Zn, Cd and Mn and Sum III>HPEM>HPE for Cr and Fe. In the case Cr, Sum II is lower than HPEM and higher than HPE. In case of Cu, extracted heavy metals is large in the order Sum IV>HPEM>Sum III HPE. HPE/HPEM value decreases with increasing the amount of HCl used for maintaining 0.1 N of extraction solution. For samples with high buffer capacity, HPE/HPEM value in all elements is lower than 0.2. On the other hand, for samples with low buffer capacity, HPE/HPEM value are over 0.2 and many samples have values higher than 0.6 for Zn, Cd Mn and Cu due to the small difference between Sum II and Sum III, and relatively higher mobility. However, for Fe and Cr, HPE/HPEM value is below 0.2 even for samples with low buffer capacity due to their low mobility and big difference between Sum II and Sum III. This study indicates that the partial extraction method in Korean Standard Method of soil is not suitable for an assessment of soil contamination in area where buffer capacity of soil can be decreased or lost because of a long term exposure to environmental damage such as acidic rain.

Surface Plasmon Resonance Ellipsometry Using an Air Injection System with an Extraction of Air System (공기주입 장치와 공기제거 장치를 사용한 표면 플라즈몬 공명 타원계측기)

  • Lee, Hong-Won;Cho, Eun-Kyoung;Jo, Jae-Heung;Won, Jong-Myoung;Shin, Gi-Ryang;CheGal, Won;Cho, Yong-Jai;Cho, Hyun-Mo
    • Korean Journal of Optics and Photonics
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    • v.20 no.3
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    • pp.182-188
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    • 2009
  • The surface plasmon resonance ellipsometer (SPRE), using a multiple air injection system with an extraction of air system, has been proposed and developed to minimize measurement error of signals due to diffusion of reagent into running buffer. Since the diffusion of reagent into running buffer affects the refractive index of the running buffer by changing the concentration, characteristics of binding between various bio-molecules don't appear clearly in measurement results. The diffusion between running buffer and reagent can be blocked by using an air bubble injection system. An extraction of air system is used to remove the noise signal due to unnecessary air bubbles flowing in a channel. Reliability of measurement results has been improved by using the valve system.

Fractionation and Electrophoretic Patterns of Rice Proteins (쌀단백질의 분획 및 전기영동)

  • Kim, Su-Il;Jo, Do-Hyun
    • Applied Biological Chemistry
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    • v.26 no.1
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    • pp.65-72
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    • 1983
  • The composition of four rice protein groups is greatly affected by the extraction conditions. The extraction amounts of albumins and glubulines primarily depended on the temperature rather than the method of extraction. The total amount of glutelins, the major components of rice storage proteins, could be extracted by a successive extraction processes, extraction with 0.5% SDS-0.1M borate buffer(pH 8.3) followed by extraction with 0.5% SDS-0.6% ${\beta}-mercaptoethanol-0.1M$ borate buffer(pH 8.3). The extracted amounts of glutelin with these solvents were 54.1 and 45% respectively. The further purification of SDS soluble glutelins was achieved by Sephadex G-150 gel column chromatography. The molecular weight of the components in four protein groups has been estimated by SDS-polyacrylamide gel electrophoresis with or without ${\beta}-mercaptoethanol.$ The comparison of albumins and globulins by starch gel electrophoresis at pH 3.1 permitted us to identify seven rice varieties. However, at pH 8.95, the specific bands for Japonica type rice varieties were observed.

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Direct Extraction of DNA from Soil for Amplification of 16S rRNA Gene Sequences by Polymerase Chain Reaction

  • Cho, Jae-Chang;Lee, Dong-Hun;Cheol, Cho-Young;Cho, Jang-Cheon;Kim, Sang-Jong
    • Journal of Microbiology
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    • v.34 no.3
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    • pp.229-235
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    • 1996
  • Microgram quantities of DNA per gram soil were recovered with SDS- based and freeze-and thaw procedures. The average DNA fragment size was > 23 Kb. This method generated minimal shearing of extracted DNA. However, the DNA extracts still contained considerable amounts of humic impurities sufficient to inhibit PCR. Several approaches were used to reduce the interferences with the PCR (use of CTAF in extraction step, Elutip-d column purification, addition of BSA to PCR buffer) to accomplish PCR with DNA extract as a template. Most of the DNA extracts were not digested completely by restriction endonuclease, and CTAB-TREATED ane Elutip-d column purified DNA extracts were partially digested. Regarding as restriction enzyme digestion, all PCRs failed to amplify 16S rRNA gene fragments in the DNA extracts. In the case of DNA extracts only where BSA was added to PCR buffer, PCR was successfully conducted whether the DNA extracts were treated with CTAB or purified with columns. However, these two treatments were indispensable for humic impurity-rich DNA extracts to generate the PCR-compatible DNA samples. Direct extraction of DNA, coupled with these procedures to remove and relieve interferences by humic impurities and followed by the PCR, can be rapid and simple method for molecular microbiological study on soil microorganisms.

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About Chromium (VI) Extraction from Fertilizers and Soils

  • Sager Manfred
    • Economic and Environmental Geology
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    • v.38 no.6 s.175
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    • pp.657-662
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    • 2005
  • Extractions fro fertilizer and soil samples were performed to yield the operationally defined fractions 'soluble' chromate (extractable with $NH_4NO_3$), 'exchangeable' chromate (extractable with phosphate buffer pH 7.2), and these results were compared with the data obtained by extractions with ammonium sulfate, borate buffer pH 7.2, saturated borax pH 9.6, and polyphosphate (Graham's salt). In order to maintain the pH of extractant solution about constant, the concentration of extractant buffer had to be raised to at least 0.5 M. The results strongly depended on the kind of extractant, and the solid: liquid ratio. For most of the samples investigated, the extraction efficiency increased in the order borate-sulfate-nitrate-phosphate. Whereas the recovery of $K_2CrO_4\;and\;CaCrO_4$ added to the samples of basic slags prior to the extraction was about complete, the recovery of added $PbCrO_4$ was highly variable. In soil extracts, the color reaction was interfered from co-extracted humics, which react with the chromate in weak acid solution during the time period necessary for color reaction (1 hour). However, this problem can be overcome by standard addition and subtraction of the color of the extractant solution. In soil extract of about pH < 7, organic material reduced chromate during the extraction period also, and standard addition of soluble chromate is recommended to prove recovery and the stability of chromate in the samples. In admixtures of soils and basic slags, results for hexavalent chromium were lower than from the mere basic slags. This effect was more pronounced in phosphate than in nitrate extracts. As a proficiency test, samples low in organic carbon from contaminated sites in Hungary were tested. The results from $NH_4NO_3$ extracts satisfactorily matched the results of the Hungarian labs obtained from $CalCl_2$ extractants.

Extraction of Changed Pixels for Hyperion Hyperspectral Images Using Range Average Based Buffer Zone Concept (구간평균 그래프 기반의 버퍼존 개념을 적용한 Hyperion 초분광영상의 변화화소 추출)

  • Kim, Dae-Sung;Pyen, Mu-Wook
    • Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
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    • v.29 no.5
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    • pp.487-496
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    • 2011
  • This study is aimed to perform more reliable unsupervised change detection through the re-extraction of the changed pixels which were extracted with global thresholding by applying buffer zone concept. First, three buffer zone was divided on the basis of the thresholding value which was determined using range average and the maximum distance point from a straight line. We re-extracted the changed pixels by performing unsupervised classification for buffer zone II which consists of changed pixels and unchanged pixels. The proposed method was implemented in Hyperion hyperspectral images and evaluated comparing to the existing global thresholding method. The experimental results demonstrated that the proposed method performed more accuracy change detection for vegetation area even if extracted slightly more changed pixels.

A Study on On-line Recognition System of Korean Characters (온라인 한글자소 인식시스템의 구성에 관한 연구)

  • Choi, Seok;Kim, Gil-Jung;Huh, Man-Tak;Lee, Jong-Hyeok;Nam, Ki-Gon;Yoon, Tae-Hoon;Kim, Jae-Chang;Lee, Ryang-Seong
    • Journal of the Korean Institute of Telematics and Electronics B
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    • v.30B no.9
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    • pp.94-105
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    • 1993
  • In this paper propose a Koaren character recognition system using a neural network is proposed. This system is a multilayer neural network based on the masking field model which consists of a input layer, four feature extraction layers which extracts type, direction, stroke, and connection features, and an output layer which gives us recognized character codes. First, 4x4 subpatterns of an NxN character pattern stored in the input buffer are applied into the feature extraction layers sequentially. Then, each of feature extraction layers extracts sequentially features such as type, direction, stroke, and connection, respectively. Type features for direction and connection are extracted by the type feature extraction layer, direction features for stroke by the direction feature extraction layer and stroke and connection features for stroke by the direction feature extraction layer and stroke and connection features for the recongnition of character by the stroke and the connection feature extractions layers, respectively. The stroke and connection features are saved in the sequential buffer layer sequentially and using these features the characters are recognized in the output layer. The recognition results of this system by tests with 8 single consonants and 6 single vowels are promising.

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Simultaneous Determination of Tin, Nickel, Lead, Cadmium and Mercury in Cigarette Material by Solid Phase Extraction and HPLC

  • Hu, Qun;Yang, Guangyu;Ma, Jing;Liu, Jikai
    • Bulletin of the Korean Chemical Society
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    • v.24 no.10
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    • pp.1433-1436
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    • 2003
  • A new method for the simultaneous determination of heavy metal ions in cigarette material by microwave digestion and reversed-phase high-performance liquid chromatography (RP-HPLC) has been developed. The cigarette material was digested by microwave digestion. Lead, cadmium, mercury, nickel and tin ions in the digested samples were pre-column derivatized with tetra-(2-chlorophenyl)-porphyrin ($T_2$-CPP) to form color chelates, which were then enriched by solid phase extraction with a $C_{18}$ cartridge. The chelates were separated on a Waters Xterra$^{TM}RP_{18}$ column by gradient elution with methanol (containing 0.05 mol/L pyrrolidine-aceticacid buffer salt, pH = 10.0) and acetone (containin0.05 mol/L pyrrolidine-acetic acid buffer salt, pH = 10.0)as mobile phase at a flow rate of 0.5mL/min and analyzed with a photodiode array detector from 350-600 nm. The detection limits of lead, cadmium, mercury, nickel and tin were 4,3,3,8 and 5 ng/L, respectively, in the original samples. This method was afforded good results.