• Title/Summary/Keyword: 2-gel-electrophoresis

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Purification of Peptide Components including Melittin from Bee Venom using gel filtration chromatography and propionic acid/urea polyacrylamide gel electrophoresis (Gel filtration chromatography와 propionic acid/urea polyacrylamide gel electrophoresis를 이용한 봉독 성분의 분리)

  • Choi, Young-Chon;Choi, Suk-Ho;Kwon, Ki-Rok
    • Journal of Pharmacopuncture
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    • v.9 no.2
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    • pp.105-111
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    • 2006
  • Objectives : This study was conducted to carry out Purification of Melittin and other peptide components from Bee Venom using gel filtration chromatography and propionic acid/urea polyacrylamide gel electrophoresis Methods : Melittin and other peptide components were separated from bee venom by using gel filtration chromatography on Sephadex G-50 column in 0.05M ammonium acetate buffer. Results : Melittin and other peptide components were separated from bee venom by using gel filtration chromatography on Sephadex G-50 column in 0.05M ammonium acetate buffer. The fractions obtained from gel filtration chromatography was analyzed by using SDS-PAGE and propionic acid/urea polyacrylamide gel electrophoresis. The melittin obtained from the gel filtration contained residual amount of phospholipase $A_2$ and a protein with molecular weight of 6,000. The contaminating proteins were removed by the second gel filtration chromatography. Conclusion : Gel filtration chromatography and propionic acid/urea polyacrylamide gel electrophoresis are useful to separate peptide components including melittin from bee venom.

Separation of large DNA molecules by pulsed field gel electrophoresis (역전장 전기영동장치를 이용한 대형 DNA 분리에 관한 연구)

  • Joo, Yi-seok;Casey, Thomas A;Yoon, Yong-dhuk
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.81-85
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    • 1993
  • Gel electrophoresis has proven to be one of the most useful of DNA separation and purification. The new technique of pulsed field gel electrophoresis (PFGE) is high resolution separation of large size DNA moleculs. Conventional continuous gel electrophresis can not be separation of large DNA fragments(20~50 k base). Field inversion gel electrophoresis(FIGE) is very useful for large DNA molecules. We have found that a pulse ratio ; 2 : 1, time ; 24hrs., volts ; $10^{volts}/_{cm}$, start ; 0.45sec, end ; 1sec, is most effectively resolves DNA fragment in the 6~50k base.

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Appropriate Electrophoresis Techniques and Isozymes to Identification of Barley Cultivars (보리품종 구분에 적합한 전기영동법과 효소)

  • Son, Eung-Ryong;Lee, Yong-Se;Yoon, Kyung-Eun;Ha, Yong-Woong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.30 no.4
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    • pp.405-411
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    • 1985
  • The buffer soluble proteins were extracted from six cultivars of barley grains and analyzed by various electrophoresis; 7.5% polyacrylamide slab gel, 2-30% polyacrylamide porosity gradient tube gel, isoelectric focusing (pH4-9) and starch gel electrophoresis. The proteins, esterase, acid phosphatase, malate dehydrogenase, glutamate dehydrogenase and leucine aminopeptidase were investigated to find out the best method to differentiate barley cultivars. The result were that protein and esterase bands in 2-30% polyacrylamide porosity gradient tube gel electrophoresis and protein bands in 7.5% polyacrylamide slab gel electrophoresis showed typical varietal differences. Therefore, those methods were suitable for differentiation of barley cultivars. It was difficult to differentiate the cultivars by the other methodes and patterns of the other enzymes.

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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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Electrophoretic Analysis of Haemolymph Proteins during Silkworm (Bombyx mori L.) Ontogenesis

  • Staykova, Teodora
    • International Journal of Industrial Entomology and Biomaterials
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    • v.14 no.1
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    • pp.37-44
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    • 2007
  • A study was made of the haemolymph protein spectrum of mulberry silkworm (Bomhyx mori L.) from the first larval instar to imago. Horizontal starch gel electrophoresis was used. Sixteen races and eight F1 interracial hybrids, raised in Bulgaria, were analyzed. During the ontogenesis, a total of 17 protein bands (15 cathodic and 2 anodic) were detected. Distinct dynamics in the haemolymph protein spectrum was observed, in result of different expression during the individual development associated with the processes of growth, histolysis and histogenesis. Based on the ontogenetic dynamics found, a correspondence was assumed between some proteins detected by us using the starch gel electrophoresis and major haemolymph proteins (SP1, SP2, MHPs and Vg) detected by other authors using the polyacrilamide gel electrophoresis. Intraracial and interracial polymorphism was observed in four protein zones. The effect of four polymorphic loci with codominant and null alleles was suggested.

Studies on Xanthine Oxidase from Bovine Thyroid Glands -[Part 1] Purification and Substrate Specificity- (소의 갑상선에 있는 크산친 옥시다아제에 관한 연구 -[제1보] 효소의 정제와 기질특이성-)

  • Lee, Hyo-Sa
    • Applied Biological Chemistry
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    • v.21 no.2
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    • pp.112-118
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    • 1978
  • Xanthine oxidase from bovine thyroid glands was purified to apparent homogeneity when judged by analytical disc gel electrophoresis. The purification procedures include pancreatin digestion, butanol extraction, ammonium sulfate precipitation, calcium phosphate gel adsorption, ultrafiltration, calcium phosphate gel-cellulose column chromatography, gel filtration, preparative Sephadex G-25 column electrophoresis, and preparative polyacrylamide gel electrophoresis. The enzyme was enriched 1,000-fold. However, its specific activity was markedly low as compared with highly purified milk enzyme. Thyroidal xanthine oxidase exhibited a low specificity for substrates and electron acceptors. The kinetic properties of thyroid xanthine oxidase were found to be similar to those of the milk enzyme on the basis of Michaelis constants for common substrates.

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Studies on the Identification of Turfgrass by Electrophoresis (SDS-PAGE, PAGIF) (전기영동법 (SDS-PAGE, PAGIF)에 의한 잔디 분류에 관한 연구)

  • 박재복;김영후;이수영
    • Asian Journal of Turfgrass Science
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    • v.5 no.1
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    • pp.11-22
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    • 1991
  • This experiment was executed to investigate the possibility of the application of taxonomic method through the isoelectric focusing with polyacrylamide gel and sodium dodecyl sulfate-polyacrylamide gel electrophoresis with seeds in the identification of turfgrasses. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to investigate the pattern of seed proteins which were extracted from 18 cultivars of cool season turfgrass and 4 cultivars of warm season turfgrass. The isoelectric focusing with polyacrylarnide gel was used to investigate the activity of the three isozymes of esterase, peroxidase and phosphoglucose isomerase which were extracted from 18 cultivars of cool season turfgrass and 4 cultivars of warm season turfgrass. The results were summarized as follows. 1. The difference of the patterns of seed proteins was observed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The identification of intra-genus was easily detected. 2. The three isozymes of esterase, peroxidase and phosphoglucose isomerase were investigated through isoelectric focusing with polyacrylamide gel. As a result, esterase was most effective among three isozymes in the identification of turfgrass cultivars 3. In the past cultivar identification was primarily based on visual morphological characters, but there was a lot of difficulty. If we should use electrophoresis, we will be able to identifvturfgrass cultivars more effectively.

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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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