• Title/Summary/Keyword: Polyacrylamide gel electrophoresis

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Differential Induction of Pathogenesis-Related Proteins in the Compatible and Incompatible Interactions of Tomato Leaves with Xanthomonas campestris pv. vesicatoria (Xanthomonas campestris pv. vesicatoria와 토마토잎의 친화적, 불친화적 반응에서 병생성관련 단백질의 유도)

  • 김정동;황병국
    • Korean Journal Plant Pathology
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    • v.11 no.1
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    • pp.53-60
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    • 1995
  • Inoculation with the compatible strain Ds 1 of Xanthomonas campestris pv. vesicatoria caused brownish ad water-soaked lesions, but incompatible strain Bv5-4a produced hypersensitive symptoms with local necrosis on tomato (cv. Kwangyang) leaves. Bacterial populations of the compatible strains Ds 1 propagated more greatly than the incompatible strain Bv5-4a at the frist onset, but no differences were observed 5 days after inoculation. The bacterial infection induced the synthesis and accumulation of soluble proteins in tomato leaves, especially in the incompatible interaction. Native-polyacrylamide gel electrophoresis distinguished the soluble proteins in the tomato leaves infected by the compatible or incompatible strains. A protein of low molecular weight occurred only in the incompatible interaction. Some pathogenesis-related (PR) proteins, especially the 15, 18, 23, 26 and 54 kDa proteins, were detected only in the infected tomato leaves. In the two-dimensional electrophoresis, some proteins with different molecular weights (Mr. 21∼29 kDa) and the pI 8∼9 appeared more distinctly only in the incompatible interaction. These data suggest that the de novo synthesis of some PR proteins in tomato may be significant in defense against X. c. pv. vesicatoria.

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Chemical Quality Evaluation of Commercial Infant Formulas (시판 조제 분유의 화학적 품질 평가)

  • 한정은;홍윤호
    • Food Science of Animal Resources
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    • v.21 no.1
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    • pp.56-63
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    • 2001
  • In this study, three domestic and one foreign formulas for the infants up to 5 month old were examined to detect chemical changes such as pH, reactive sulfhydryl groups(RSH) content, 5-hydroxymethylfurfral(HMF) content, available lysine content, electrophoresis, and surface color caused by heat treatment for long term storage. In the SDS-polyacrylamide gel electrophoresis, A and B products showed similar pattern, while C product had a clearly distinguishable $\beta$-lacto-globulin band, but in casein, only D product showed a few strong casein band. RSH content, which indicate the extent of whey protein denaturation, ranged from 4.40 to 5.93 mmole/g protein. HMF content. which indicate the extent of Maillard reaction, ranged from 192 to 432 $\mu$mole/100g in formulas. B product showed the highest RSH and HMF content. Available lysine content ranged from 31 to 46 mg/g protein. Among them D product contain the highest available lysine content and others showed no significant difference. In conclusion, the domestic infant formulas showed higher RSH and HMF content than the foreign product and the available lysine content of the domestic products were lower than of the foreign product.

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Electrophoretic Studies on Haemolymph Protein, Digestive Fluid Protein and Digestive Amylase Activity in the Wild silkworm, Theophila mandarina. (상잠의 형액과 소화액에 있어서 단백질과 Amylase 활성에 관한 전기영동적 연구)

  • 문재유;박광의
    • Journal of Sericultural and Entomological Science
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    • v.27 no.1
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    • pp.37-41
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    • 1985
  • The haemolymph protein, digestive fluid proteins and digestive fluid amylase activity of wild silkworm, Theophila mandarina those of the were studied by polyacrylamide gel electrophoresis. In addition, they was also compared with silkworm. 1. 6 main protein bands in female and 7 main protein bands in male were detected in the larval haemolymph of T. mandarina where as 8 and 7 main protein bands in female and male of B. mori were observed. Some differences in the haemolymph protein ands of T. mandarina and B. mori were observed. 2. 15 protein bands and 12 protein bands were found in the larval digestive fluid of T. mandarina and B. mori respectively. Some differences in the mobility of digestive fluid proteins of T. mandarina and B. mori were noticed. 3. Larval digestive fluid amylases were anionic and moved near the tracking dye in both T. mandarina and B. mori. Mobility of the digestive fluid amylases relative to bromophenol blue were 0.019 and 0.020 in T. mandarina and B. mori respectively.

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Characterization of a Toxin isolated from Bacillus thuringiensis var. alesti and Its Toxicity to the Silkworm, Bombyx mori (Bacillus thuringiensis var. alesti로부터 분리된 독소의 성상과 누에에 대한 독성)

  • 이영근;조용섭;임종성
    • Journal of Sericultural and Entomological Science
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    • v.17 no.1
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    • pp.55-62
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    • 1975
  • In the studies of characterization of crystal prototoxin isolated from Bacillus thuringinensis var. alesti and its bioassay to the silkworm larvae, Bombyx mori L., the following results were obtained. 1. The complete lysis of bacteria grown on the nutrient agar at 30$^{\circ}C$ in an incubator took 30 days, but the period could be reduced by a half in a devised broth media in a fermentation jar. 2. The protein toxin extracted directly from a mixture of crystals and spores without separation of crystals and spores was pure as same as the protein toxin extracted from only crysals separated. 3. By SLS polyacrylamide gel electrophoresis of prototoxin released from crystals in alkali (pH 9.5) three different proteins in molecular weights of 120,000, 87,000, 74,000 were separated. 4. In the bioassay of the toxin to the silkworm larvae, LD$\sub$50/ per gram of the larvae in the 4th instar was 0.080 $\mu\textrm{g}$.

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Purification and Properties of Alkaline Pretense from Xanthomonas sp. YL-37 (Xanthomonas sp. YL-37 균주가 생산하는 Alkali성 단백질분해효소의 정제 및 성질)

  • 장형수;권태종
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.427-434
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    • 1998
  • An alkaline protease was 4-fold purified, yielding 2.3% of recovery by ammonium sulfate precipitation, CM-cellulose column chromatography and Sephadex G-100 column chromatography. The purified enzyme was estimated to be monomeric with molecular weight of about 62,000 from polyacrylamide gel eletrophoresis (PAGE) and sodiumdodecylsulfate polyacrylamide gel electrophoresis (SDS-FAGE). The optimal pH and temperature of the alkaline pretense activity were 11.0 and 50$^{\circ}C$, respectively, exhibiting high stability at pH value from 6.0 to 11.0 at 50$^{\circ}C$ for 30 minute. The alkaline pretense was activated by MnSO$_4$, CaCl$_2$, and was inhibited by CuSO$_4$, ZnSO$_4$, HgCl$_2$, EDTA and EGTA. Also, the enzyme was found to be a metaloenzyme requiring Mn$\^$2+/ as cofactor. The NH$_2$-terminal amino acid of alkaline protease was alanine. The Km and Vmax values of this enzyme for casein was 4.0 mg/$m\ell$ and 5,500 unit/$m\ell$, respectively.

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Isolation of Iron and Calcium-Binding Peptides from Cottonseed Meal Protein Hydrolysates (면실박 단백질로부터 가수분해물 제조 및 철분, 칼슘 결합 펩타이드의 분리)

  • Choi, Dong-Won;Kim, Nam-Ho;Song, Kyung Bin
    • Journal of Applied Biological Chemistry
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    • v.55 no.4
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    • pp.263-266
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    • 2012
  • Isolation of iron and calcium-binding peptides derived from cottonseed meal protein (CMP) hydrolysates was investigated. The degree of hydrolysis of CMP by Flavourzyme was monitored using trinitrobenzenesulfonic acid method and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Enzymatic hydrolysis of CMP for 12 h was sufficient for the preparation of CMP hydrolysates, and the hydrolysates were membrane-filtered under 3 kDa as a molecular weight. The filtered solution was fractionated using Q-Sepharose fast flow, Sephadex G-15, and reversed phase-high performance liquid chromatography for iron and calcium-binding peptides. As a result, F51 fraction was obtained as the best candidate for calcium and iron chelation, and the isolated iron and calcium-binding peptides can be used as functional food additives, similar to iron and calcium supplements.

Purification, Characterization of Pullulanase Produced by Aerornonas caviae No. S-76 and Synthesis of Maltosyl-$\beta$-Cyclodextrin (Aeromonas caviae No. S-76이 생산하는 Pullulanase의 정제, 특성 및 Maltosyl-$\beta$-Cyclodextrin의 합성)

  • 손천배;김명희;이명자
    • Microbiology and Biotechnology Letters
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    • v.19 no.4
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    • pp.362-367
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    • 1991
  • The crude enzyme solution obtained by shaking culture of Aeromonas caviae No. S-76 isolated from soil as pullulanase producing bacterium was purified by 50 folds with 21% yield by salting out with ammonium sulfate and column chromatography using DEAE-Sephadex A-50 and Sephadex G-150. The purified pullulanase had a molecular weight of 118, 000 approximately by SDS-polyacrylamide slab gel electrophoresis and pI of 4.3 by isoelectric focusing. And optimum reaction temperature and pH for puHulanase were $50^{\circ}C$ and 8.0, respectively. The purified enzyme was relatively stable at pH 6.0~9.0 and below $45^{\circ}C$. This enzyme synthesized maltosyl-$\beta$-cyclodextrin from mixture of $\beta$-cyclodextrin and maltose.

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Purification and Immunological Properties of Vitellin, and its Quantitative Changes during Embyrogenesis in the silkworm, Bombyx mori (누에 난황소(Vitellin)의 분리와 면역학적 특성 및 발육에 따른 함량변화)

  • 손기국;문재유
    • Journal of Sericultural and Entomological Science
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    • v.30 no.2
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    • pp.96-104
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    • 1988
  • Vitellin, the major yolk protein of the silkworm, Bombyx mori was pruified, and its immunological properties and the quantitative changes during embryogenesis were studied. The ovary transplantation into male hosts was also carried out to find its effect on the yolk protein synthesis. The pupal vitellogenin and the egg vitellin of Bombyx mori were purified by DEAE-cellulose column chromatography. These two female specific proteins showed the same mobility in the polyacrylamide gel electrophoresis and the same reaction in the double immunodiffusion test. The immunological identity was also observed between the vitellins of Bombyx mori and Bombyx mandarina. The rudimentary ovaries transplanted into the male hosts of silkworms produced eggs without vitellin, indicating that the yolk precursors synthesized in other female organ beyond the ovary were necessary to produce vitellins. The major yolk protein, vitellin was disintegrated and utilized mostly during late stage of embryogenesis. It was different characteristics from the egg specific protein, which was utilized continuously from the early embryonic stage.

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Purification of Glucose Oxidase by Affinity Chromatography and Its Characterization (친화성 크로마토그래피를 이용한 글루코오스 옥시다아제의 정제와 효소특성)

  • Ko Jung Hwan;Byun Si Myung
    • Journal of the Korean Chemical Society
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    • v.23 no.3
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    • pp.165-174
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    • 1979
  • A purification technique of glucose oxidase was developed. Using the gluconyl-${\omega}$-aminohexyl Sepharose affinity chromatography, it was partially purified 14.6 folds with 79.7% yield. With the combination of the affinity chromatography and Sepharose 6B gel filtration, the enzyme was purified 27.2 folds from the broth with 74.1% yield. The final purified preparation showed 90.83 U of glucose oxidase activity per mg of protein and a single band by 7% polyacrylamide gel electrophoresis. The absorption spectrum and substrate specificity of the enzyme were studied and the fianal preparation showed the optimal pH between 5.6 and 6.0, the optimal temperature at $40^{\circ}C$, $8.5{\times}10^{-3}M$ of $K_m$ for D-glucose, and 3.43 kcal/mole of the activation energy.

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Trypsin Inhibitor Variants in Korean Land Races and Wild Soybeans (한국 재래종 및 야생종 대두의 Trypsion Inhibitor 변이)

  • Shin-Han Kwon;Mi Ryung, Chae;Kyung Sook, Park;Hi Sup, Song
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.35 no.2
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    • pp.171-175
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    • 1990
  • One thousand and seven hundreds and six soybean land reces and 103 wild soybeans of Korea, and 167 cultivars and 71 wild soybeans of exotic were analyzed using polyacrylamide gel electrophoresis to study the distribution of trypsin inhibitor phenotypes. The four ti/ti lines were observed only in Korean native land races. The Ti$\^$*/a allele of cultivar was found to be the highest in Chinese soybean (0,9888) than in Korean soybean(0.8347) and Japanese soybean (0.5954). The heterozygosity of typsin inhibitor in Korean lines occured in relatively high ratio, The percentages of heterozygosity of the Korean land race and wild soybean are 3.6% (N=61) and 9.7%(N=10), respectively.

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