• Title/Summary/Keyword: Gel electrophoresis

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Studies on Xanthine Oxidase from Bovine Thyroid Glands -[Part 2] Composition and Some Properties- (소의 갑상선에 있는 크산친 옥시다아제에 관한연구 [제2보] 효소의 조성과 특성-)

  • Lee, Hyo-Sa
    • Applied Biological Chemistry
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    • v.21 no.3
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    • pp.137-143
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    • 1978
  • Xanthine oxidase from bovine thyroid glands was found to contain FAD, molybdenum and iron in a ratio 1:0. 36:1. 6. The molecular weight of the thyroid enzyme was similar to that of the milk enzyme when estimated by gel filtration and polyacrylamide gel electrophoresis. The optimum pH for the enzyme activity was 7.8. The pH of the isoelectric point was determined to be 6.2 by electrofocusing. Sodium dodecyl sulfatepolyacrylamide gel electrophoresis experiment indicated that the enzyme was dissociated into subunits and that the molecular weight for the smallest subunit was 65,000 daltons. Absorption spectra were dissimilar between milk and thyroid xanthine oxidase.

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A rapid method for sodium dodecyl sulfate polyacrylamide gel electrophoresis evaluation (Sodium dodecyl sulfate polyacrylamide gel 전기영동 결과의 경제적인 평가 방법)

  • Page, C.;Hamelin, C.
    • Korean Journal of Microbiology
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    • v.27 no.4
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    • pp.436-438
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    • 1989
  • We propose a method for rapid evaluation and permanent record of sodium dodecyl sulfate polyacrylamide gel electrophoretic runs. This method is based on the photocopy process, rather than on photography and requires no extensive or expensive investment. Comparison of a print obtained through this method and a 35mm photography indicated, on a balanced gel, equal sensitivity.

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Proteins and Amino Acid Composition of Korea Ginseng Classified by Years (한국인삼의 연근별 단백질 및 아미노산 조성)

  • Choi, Cheong;Yoon, Sang-Hong;Bae, Man-Jong;An, Bong-Jean
    • Korean Journal of Food Science and Technology
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    • v.17 no.1
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    • pp.1-4
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    • 1985
  • For the systematic investigation of biochemical characteristics of Korean ginseng protein by years, protein fractions were analyzed by the techniques of polyacrylamide gel electrophoresis, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration, while the amino acid composition was studied by amino acid autoanalyzer. Results of polyacrylamide gel electrophoresis and SDS-PAGE showed a few difference in pattern and number of bands depending on the age of the root. However, the number of bands obtained from polyacrylamide gel electrophoresis and SDS-PAGE was 8 and 7 to 11, respectively. When water extracted proteins were fractionated by Sephadex G-200, the main peak among 2 peaks was collected and lyophillized. Its mol. wt. was extimated to be 43,000 by the SDS-PAGE method. In amino acid composition of water extracted protein and main fraction of gel filtration, arginine content was the highest, 47.17% in water extracted protein and 57.36% in main fraction followed by glutamic acid and asparatic acid. On the contrary, cystine and methionine contents were very low in both cases.

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A Study of Alcohol Dehydrogenase of Drosophila melanogaster in Korea by Starch Gel Electrophoresis (Starch gel 전기영동법에 의한 한국산 초파리의 alcohol dehydrogenase (ADH)에 관한 연구)

  • Chung, Yong-Jai
    • The Korean Journal of Zoology
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    • v.16 no.1
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    • pp.67-77
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    • 1973
  • The alcohol dehydrogenase (ADH) isozyme of the four strains of Drosophila melanogaster in Korea was studied by the starch gel electrophoresis and the results are given below: 1. The numbers of bands, staining intensity and mobility are considerably variable in the three strains, Chunchun, Jinju and Sinchon-Seoul, but relatively uniform in the Taijun strain. 2. It is suggested that the Taijun strain may be homogeneous and the other three strains, on the contrary, heterogeneous for the ADH constitution. 3. The electrophoretic patterns are observed to be different between males and females in all strains used which is hard to intereret. 4. The starch gel electrophoresis is to have a resolving power superior to cellulose acetate electrophoresis.

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Bacterial Community Analysis during Composting of Garbage using Denaturing Gradient Gel Electro-phoresis (Denaturing Gradient Gel Electrophoresis를 이용한 음식물 쓰레기 퇴비화 세균 군집 분석)

  • Ryu Hee Wook;Cho Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.33 no.3
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    • pp.226-230
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    • 2005
  • The microbial community during composting of gargage was analyzed using 16S rDNA PCR - DGCE (denaturing gradient gel electrophoresis). Pseudomonas spp. was found throughout the process, and thermophilic Bacillus spp. was dominated at the thermophilic stage. Six thermophilic bacteria were isolated and identified as B. caldoxylolyticus, B. thermoalkalophilus, and B. thermodenitrificans.

Single Cell Gel Electrophoresis (comet assay) to Detect DNA Damage and Apoptosis in Cell Level (DNA damage와 Apoptosis를 정량화하는 단세포전기영동법)

  • 류재천;김현주;서영록;김경란
    • Environmental Mutagens and Carcinogens
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    • v.17 no.2
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    • pp.71-77
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    • 1997
  • The single cell gel electrophoressis(SCGE) assay, also known as the comet assay, is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakage in mammalian cells. The SCGE or comet assay is a promising test for the detection of DNA damage and repair in individnal cells. It has widespread potential applications in DNA damage and repair studies, genotoxicity testing and biomonitoring. In this microgel electrophoresis technique, cells are embedded in agarose gel on microscope slides, iysed and electrophoresed under alkaline conditions. Cells with increased DNA damage display increased migration of DNA from the nucleus towards the anode. The length of DNA migration indicates the amount of DNA breakage in the cell. The comet assay is also capable of identifying apoptotic cells which contain highly fragmented DNA. Here we review the development of the SCGE assay, existing protocols for the detection and analysis of comets, the relevant underlying principles determining the behaviour of DNA and the potential applications of the technique.

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A Preliminary Analysis of Secreted Proteins from Bifidobacterium pseudocatanulatum BP1 by Two-Dimensional Gel Electrophoresis

  • Moon, Gi-Seong
    • Preventive Nutrition and Food Science
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    • v.13 no.4
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    • pp.366-369
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    • 2008
  • Proteins secreted from bifidobacteria are believed to play important roles in human intestines via interacting with different host cells. In this respect, proteins secreted from Bifidobacterium pseudocatanulatum BP1, which has been rarely studied, were analyzed by two-dimensional gel electrophoresis (2DE). Using this approach, approx-imately 21 protein spots on a 2DE gel were detected and 10 of these spots were identified by mass spectrometry. Five spots were identified as hypothetical proteins and the remaining 5 spots were identified as a putative iron-side-rophore binding lipoprotein, a short-chain dehydrogenase/reductase SDR, an exonuclease, cytochrome P450 hydroxylase, and a putative dehydrogenase. The identification of secreted putative iron-siderophore binding lipoprotein was highly interesting since it is an important protein that is involved in ferric iron uptake in pathogenic bacteria. This finding could accelerate studies on the probiotic effect of Bifidobacterium by explaining the competition between bifidobacteria and intestinal pathogens for ferric iron.

Purification and Properties of .$\beta$-1, 3-Glucanase from Pseudomonas stutzeri KF13 (Pseudomonas-stutzeri KF13의 ..$\beta$-1, 3-Glucanase 정제 및 성질)

  • 방광웅;송형익;김재근;유대식;정기택
    • Korean Journal of Microbiology
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    • v.25 no.1
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    • pp.1-8
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    • 1987
  • An extracellular $\beta$-1, 3-glucanase from Pseudomonas stutzeri KF 13 was purified about 390 with 26% recovery. The purified enzyme revealed a single band by polyacrylamide gel electrophoresis and SDS-polyacrylamide gel electrophoresis. The enzyme was stable in a pH 6.0 to 9.0, and relatively thermostable. The optimal pH and temperature on the enzyme activity were found to be 5.8 and 45.deg.C, respectively. The activation energy was calculated to be 16,130 cal per mole. The Km value for laminarin was found to be 3ng per ml and the molecular weight was determined to be 28,000 by gel filtration and 26,000 daltons by SDS-acrylamide gel electrophoresis. The enzyme was inhibited by 1.0mM of $Hg^{2+}$, and strongly inhibited by 1.0mM of p-chloromercuribenzoic acid.

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Protein Separation in Functional Rice Grains Using Two-Dimensional Gel Electrophoresis

  • Chung, Soo Im;Lee, Sang Chul;Kan, Mi Young
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.58 no.4
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    • pp.388-392
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    • 2013
  • The proteins from functional rice cultivars (Nogwonchalbyeo, Giant embryonic, Arhyangchalbyeo, and Goamibyeo) and general white rice were extracted and separated using two-dimensional (2D) gel electrophoresis. A wide variation in the molecular weight (MW) and pH range of the expressed proteins in rice samples were observed. The green-kerneled rice (Nogwonchalbyeo) exhibited proteins with MW of 9-57 kDa and appeared at a pH range of 4-7. The Giant embryonic contained proteins with MW of 31-63 kDa and a pH range of 5-6. The aromatic glutinous rice (Arhyangchalbyeo) showed proteins with MW of 24-28 and pH of 5.8-6.8. The high-amylose rice (Goamibyeo) exhibited proteins with MW of 3-63 and pH of 5.2-5.6. The identified proteins uniquely found and highly expressed in each cultivar may have a significant role on rice functionality. The results illustrate that the 2D gel electrophoresis is a valuable method in the determination of the protein expression profiles in functional rice grains and may be useful in the identification of specific marker proteins associated with the functional property of rice.

Purification of Bovine Pregnancy-Associated Proteins by Two-Dimensional Gel Electrophoresis

  • Hwang, Sun-Il;Lim, Jin-Kyu
    • BMB Reports
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    • v.32 no.5
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    • pp.445-450
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    • 1999
  • We purified and characterized a bovine pregnancy-associated protein in pregnant cow urine using two-dimensional gel electrophoresis. Urine from cows was collected according to their status of pregnancy and non-pregnancy. Proteins in the cow urine were fractionated with 50% ammonium sulfate prior to two-dimensional gel electrophoresis. Proteins separated on the gels were compared in terms of expression level and new expression by molecular mass and isoelectric point. We localized two pregnancy-associated protein spots on the gels at molecular masses of 24 kDa and 20 kDa and isoelectric points of 5.5 and 5.7, respectively. Likewise, two non-pregnancy specific proteins were localized at 27 kDa and 28 kDa with isoelectric points of 5.7 and 5.9, respectively. To rule out the possibility that environmental or genetic factors might influence the expression of the proteins, we demonstrated the pregnancy-associated expression of the proteins in two-dimensional gels with pregnant urine taken from cows raised in a different institute. The pregnancy-associated protein with molecular mass of 20 kDa and isoelectric point of 5.7, namely spot 2, was microsequenced and found to be highly homologous to the bovine collagen alpha 1 chain.

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