• Title/Summary/Keyword: gel mobility

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Purification and Properties of Escherichia coli-Corynebacterium nephridii Hybrid Thioredoxin

  • Sa, Jae-Hoon;Lee, Hee-Bong;Lim, Chang-Jin
    • BMB Reports
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    • v.29 no.2
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    • pp.116-121
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    • 1996
  • In earlier studies, the genes encoding Escherichia coli thioredoxin and Corynebacterium nephridii thioredoxin C-3 were fused via a common restriction site in the nucleotide sequence coding for the active site of the proteins to generate two chimeric thioredoxins, designated E-C3 (N to C-terminal) and C3-E. The hybrid thioredoxins were overexpressed in E. coli from the cloned chimeric thioredoxin genes by a T7 promoter/polymerase system. To investigate the structure-function relationship of thioredoxin, we purified the E-C3 hybrid thioredoxin through ammonium sulfate fractionation, DEAE-cellulose chromatography, and Sephadex G-50 gel filtration. Its purity was examined on SDS-polyacrylamide gel electrophoresis and the molecular weight of the purified E-C3 hybrid thioredoxin was estimated to be 12,000. On native polyacrylamide gels, the purified E-C3 hybrid thioredoxin shows a much lower mobility than E. coli thioredoxin. E-C3 hybrid thioredoxin exhibits a 40-fold lower catalytic efficiency with E. coli thioredoxin reductase than E. coli thioredoxin. It was shown to catalyze the reduction of insulin disulfide by dithiothreitol. The purified E-C3 hybrid thioredoxin was also characterized in other aspects.

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Studies on the Purification and Biochemical Properties of Vitellin in the Antheraea yamamai Guerin-Meneville I. Isolation and Purification of Vitellin and its Change to Embryonic Development (천잠(Antheraea yamamai) Vitellin의 분리와 생화학적 특성에 관한 연구 I. Vitellin의 분리와 동정 및 배자발생에 따른 변동)

  • 김철명;문재유
    • Journal of Sericultural and Entomological Science
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    • v.31 no.2
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    • pp.72-81
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    • 1989
  • Antheraea yamamai vitellin was purified from matured eggs by polyacryamide gel electrophoresis, also stage dependent appearance, immunological comparison and relative content of the protein were investigated. 1. Vitellogenin, the precursor of vitellin, was first detected in the larval hemolymph at the late spinning stage by polyacrylamide gel electrophoresis and immunoelectrophoresis. 2. The electrophoretic mobility of the vitellin was identical with that of Bombyx mori and of Bombyx mandarina. However, the specific antiserum against A. uamamai vitellin did not react with either that of Bombyx mori or Bombyx mandarina in immumo-diffusion test. 3. Relative content of A. yamamai vitellin to the total soluble egg protein was 46.0 percent and did not change till eight days after oviposition. But the content started to decline from ten days after oviposition and was negligible in the five or serventeen month old eggs.

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A Quick Novel Method to Detect the Adulteration of Cow Milk in Goat Milk

  • Lee, Chi-Chei;Chang, His-Shan;Sheen, Hua-Shan
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.3
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    • pp.420-422
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    • 2004
  • This study was to demonstrate a rapid novel method for detection of adulterated cow milk in goat milk using modified staining protocol after native polyacrylamide gel electrophoresis (PAGE). Samples of cow milk and goat milk containing 0, 0.5, 1.0 and 2.0% (v/v) of cow milk were analyzed. Low levels of cow milk mixed in goat milk were identified by the presence of higher mobility of $\beta$-lactoglobulin A ($\beta$-Lg A) in cow milk. By mini-gel electrophoresis, a distinguishable protein profile was visualized in 25 min using the modified Coomassie blue staining solution, in which methanol (50%) was replaced with ethanol (20%) and the concentrations of Coomassie blue and acetic acid were reduced from 2 to 0.13% and 10 to 5%, respectively. To visualize the milk proteins, gels in the staining solution were water-bathed in boiling water for 5 min and then cooled down immediately for 3-5 min. The sensitivity of this method is relatively high, allowing examination of 1% cow milk in goat milk. The procedure presented here is also very cost-effective due to less reagents needed. This simplified method would be useful and applicable to dairy industry for routine examination of goat milk.

Immunological Identification of Thiobacillus ferro-oxidans and Thiobacillus thiooxidans (Thiobacillus ferrooxidans와 Thiobacillus thiooxidans의 면역학적 동정)

  • 이강순;장정순;이강석
    • Korean Journal of Microbiology
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    • v.16 no.1
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    • pp.30-40
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    • 1978
  • Detergent soluble fractions were obtained from T. ferrooxidans ATCC 13598 and the T. thiooxidans ATCC 8085 which were treated with 3% of Tween 20. The detergent soluble antigen(crude antigen) fractions of the T.ferrooxidans and the T.thiooxidans were subjected to hydroxyapatite. In the case of T.thiooxidans, further purification was carried out on the DEAE-cellulose column chromatography. The antigen fractions, such as the hydroxyapatite peak-1(Tf, HA-1) and peak-2 from T.ferrooxidans(Tf. HA-2) and hydroxyapatite peak-1(Tt, HA-1), DEAE-cellulose peak-1(Tt, DP-1) and peak-2(Tt, DP-2) from T. thiooxidans wre compared each other with the homologous and the heterologous and the heterologous antisera against to the Thiobacillus species. The hydroxyapatite peak-2 fraction from the T.ferrooxidans(Tf, HA-2) and DEAE-cellulose peak-2 fraction from the T.thiooxidans(Tt, DP-2) were represented the type-specific immuno-reactivities between the T.ferrooxidans and the T.thiooxidans on the several sets of double gel diffusioin analysis. The type-specific antigen fractions from both of the baceteria were mainly composed of protein with entierly different electrophoretic mobility on the SDS-polyacrylamide gel electrophoresis. However, the PAS positive banding patterns on the electrophorogram showed wide range of common antigenic properties in the T. ferrooxidans and the T.thiooxidans, respectively.

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Purification and Characterization of the Red Carotenoprotein from the Skin of Ascidian, Halocynthia roretzi 1. Purification and Characterization of the Caritenopritein (멍게껍질로부터 분리제정한 적색 Carotenoprotein의 특성 1. Carotenoprotein 의 정제 및 특성)

  • Kang, Ok-Ju;Suh, Myung-Ja;Lee, An-Jong;Kim, Se-Kwon
    • Journal of Life Science
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    • v.5 no.4
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    • pp.170-180
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    • 1995
  • A carotennnoprotein from the skin of Ascidian(Halocynthia roretzi) was extracted by Triton X-100 and purified by ammonium sulfate fraction, SephadexG-200 charomatography and DEAE-cellulose ion exchange chromatography. The carotenoprotein was redwith broad $\lambda$$_{max}$ between 495, 467 and 318nm. The red carotenoprotein had an approximate molecular weight of 326KDa(gel filtration). SDS-PAGE indicated the presence of two polypeptodes of 84.1KDa and 74.4KDa, with different mobility in polyacrylamide gel electrophoresis. In the presence of denaturing agents such as organic solvent aand extreme pH, the red complex readily disociates to liberate the yellow carotenoid($\lambda$$_{max}$ 452nm) and a colourless apoprotein. The amino acid composition of carotenoprotein were mainly threonine(15.2%), aspartic acid(12.2%), glutamic acid(11.9%) and serine(9.6%), while proline was not found. The carotenoprotein consisted of lipids as structure units. Its major fatty acids composion were C$_{18:1}$, C$_{16:1}$, and C$_{16:0}$. The monounsaturated fatty acids(41.5%) contained abundant content compared to other fatty aacids(polyunsaturated fatty acids 37.4%, saturated fatty acids 20.6%).

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Sol-gel deposited TiInO thin-films transistor with Ti effect

  • Kim, Jung-Hye;Son, Dae-Ho;Kim, Dae-Hwan;Kang, Jin-Kyu;Ha, Ki-Ryong
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.02a
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    • pp.200-200
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    • 2010
  • In recent times, metal oxide semiconductors thin films transistor (TFT), such as zinc and indium based oxide TFTs, have attracted considerable attention because of their several advantageous electrical and optical properties. There are many deposition methods for fabrication of ZnO-based materials such as chemical vapor deposition, RF/DC sputtering and pulsed laser deposition. However, these vacuum process require expensive equipment and result in high manufacturing costs. Also, the methods is difficult to fabricate various multicomponent oxide semiconductor. Recently, several groups report solution processed metal oxide TFTs for low cost and non vacuum process. In this study, we have newly developed solution-processed TFTs based on Ti-related multi-component transparent oxide, i. e., InTiO as the active layer. We propose new multicomponent oxide, Titanium indium oxide(TiInO), to fabricate the high performance TFT through the sol-gel method. We investigated the influence of relative compositions of Ti on the electrical properties. Indium nitrate hydrate [$In(NO^3).xH_2O$] and Titanium isobutoxide [$C_{16}H_{36}O_4Ti$] were dissolved in acetylacetone. Then monoethanolamine (MEA) and acetic acid ($CH_3COOH$) were added to the solution. The molar concentration of indium was kept as 0.1 mol concentration and the amount of Ti was varied according to weighting percent (0, 5, 10%). The complex solutions become clear and homogeneous after stirring for 24 hours. Heavily boron (p+) doped Si wafer with 100nm thermally grown $SiO_2$ serve as the gate and gate dielectric of the TFT, respectively. TiInO thin films were deposited using the sol-gel solution by the spin-coating method. After coating, the films annealed in a tube furnace at $500^{\circ}C$ for 1hour under oxygen ambient. The 5% Ti-doped InO TFT had a field-effect mobility $1.15cm^2/V{\cdot}S$, a threshold voltage of 4.73 V, an on/off current ratio grater than $10^7$, and a subthreshold slop of 0.49 V/dec. The 10% Ti-doped InO TFT had a field-effect mobility $1.03\;cm^2/V{\cdot}S$, a threshold voltage of 1.87 V, an on/off current ration grater than $10^7$, and a subthreshold slop of 0.67 V/dec.

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Theoretical Calculation on Alpha Track Density by Using an Electrostatic Ion Spectrometer (정전기분광분석방법에 의한 알파입자비적밀도의 이론적 계산연구)

  • Yoon, Suk-Chul;Ha, Chung-Woo
    • Journal of Radiation Protection and Research
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    • v.18 no.1
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    • pp.25-35
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    • 1993
  • To develop a technique of theoretical alpha track density calculation for comparison with measured track density, an electrostatic ion spectrometer was specially designed and fabricated. The mobility spectrum of first radon daughter(Po-218) in the range of $0.07{\sim}5.0cm^2/V\;s$ from the radon chamber was measured using-the electrostatic ion spectrometer. Measurement was taken in a radon chamber operated using dry particle free air passed through silica gel, activated charcoal and molecular sieve filters. The mobility of a new-born Po-218 ion measured by the electrostatic ion spectrometer was determined to be $1.92cm^2/V\;s$. A comparison of the theoretical and measured alpha track densities was completed and uncertainties concerning the shape of the spectrum were analyzed. It was found that the discrepancies in track densities are primarily Que to the neglect of wall loss of ions in the theoretical track density calculation.

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EFFECTS OF COPPER AND SALT ON THE HEMOGLOBIN OF SEVERAL FRESHWATER FISHES (담수어류 수종의 혈색소에 미치는 동과 염의 영향)

  • PARK Young Shik;LEE Choon Koo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.5 no.4
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    • pp.105-107
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    • 1972
  • The effects of copper and salt on the hemoglobin patterns of Carassius carassius, Ophicephalus argus, and Misgurnus anguillicaudatus were investigated by vertical starch gel electrophoresis. The number of the hemoglobin band of normal C. cararsius on the anode was one, and that of O. argus was two; neither the number of the homoglobin bands of the fishes nor the mobility of them was affected by copper or salt. The hemoglobin mobility of O. argus was faster than those of C. carassius and M. anguillicaudatus. Normal M. anguillicaudatus had two hemoglobin bands with nearly identical mobilities, one band migrating anodically and the other cathodically. When M. anguillicaudatus was exposed to copper, the mobility of band on the anode was faster than those of the other groups of control and exposed to salt, whereas the hemoglobin band on the cathode was not affected.

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Cloned Placenta of Korean Native Calves Died Suddenly at Two Months after Birth Displays Differential Protein Expression

  • Kim Hong Rye;Kang Jae Ku;Lee Hye Ran;Yoon Jong Taek;Seong Hwan Hoo;Jung Jin Kwan;Park Chang Sik;Jin Dong Il
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.63-68
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    • 2005
  • Cloned calves derived from somatic cell nuclear transfer (SCNT) have been frequently lost by sudden death at 1 to 3 month following healthy birth. To address whether placental anomalies are responsible for the sudden death of cloned calves, we compared protein patterns of 2 placentae derived from SCNT of Korean Native calves died suddenly at two months after birth and those of 2 normal placentae obtained from AI fetuses. Placental proteins were separated using 2-Dimensional gel electrophoresis. Approximately 800 spots were detected in placental 2-D gel stained with coomassie-blue. Then, image analysis of Malanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between normal and SCNT placentae. In the comparison of normal and SCNT samples, 8 spots were identified to be up-regulated proteins and 24 spots to be down-regulated proteins in SCNT placentae, among which proteins were high mobility group protein HMG1, apolipoprotein A-1 precursor, bactenecin 1, tropomyosin beta chain, $H^+-transporting$ ATPase, carbonic anhydrase II, peroxiredoxin 2, tyrosine-rich acidic matrix protein, serum albumin precursor and cathepsin D. These results suggested that the sudden death of cloned calves might be related to abnormal protein expression in placenta.

Electrical and optical properties of Al and F doped ZnO transparent conducting film by sol-gel method (Sol-gel법에 의한 Al과 F가 첨가된 ZnO 투명전도막의 전기 및 광학적 특성)

  • Lee, Seung-Yup;Lee, Min-Jae;Park, Byung-Ok
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.16 no.2
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    • pp.59-65
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    • 2006
  • Al-doped and F-doped ZnO (ZnO : Al & ZnO : F) thin films were coated onto glass substrate by sol-gel method. These films showed c-axis orientation in common, but different I(002)/[I(002) + I(101)] and FWHM (full width at half-maximum). In particular, the grain size of the ZnO : Al films decreased with the increase in the Al-doping concentration, while for the ZnO : F films the grain siae increased up to F 3 at% and then decreased. For the electrical properties, Hall effect measurement was used. The resistivity of the ZnO : Al films and the ZnO : F films were, respectively, $2.9{\times}10^{-2}{\Omega}cm$ at Al 1 at% and $3.3{\times}10^{-1}{\Omega}cm$ at F 3 at%. Moreover compared with ZnO:Al films, ZnO:F films have lower carrier concentration (ZnO : Al $4.8{\times}10^{18}cm^{-3}$, ZnO : F $3.9{\times}10^{16}cm^{-3}$) and higher mobility (ZnO : Al $45cm^2/Vs$, ZnO : F $495cm^2/Vs$). For average optical transmittances, ZnO : Al thin films have $86{\sim}90%$ and ZnO : F films have $77{\sim}85%$ comparatively low.