• Title/Summary/Keyword: polyacrylamide,

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Sialoglycoproteins of Mammalian Erythrocyte Membranes: A Comparative Study

  • Sharma, Savita;Gokhale, Sadashiv M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.12
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    • pp.1666-1673
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    • 2011
  • The presence of sialoglycoproteins (SGPs) in the membranes from goat (Capra aegagrus hircus), buffalo (Bubalus bubalis bubalis) and pig (Sus scrofa domestica) erythrocytes was investigated by partial purification with a chloroform-methanol extraction method followed by Sodium dodecyl sulphate - Polyacrylamide gel electrophoresis in comparison to human (Homo sapiens) erythrocytes. The results show that mammalian erythrocytes possess clear differences in the SGPs numbers and molecular weights although all animals studied in this experiment are from the same class i.e. mammalia. The SGPs number in human, goat, buffalo and pig are four (PAS-1 to PAS-4), ten (PAS-GI to PAS-GX), seven (PAS-BI to PAS-BVII) and four (PAS-PI to PAS-IV) respectively as indicated by staining the polyacrylamide gel with sialoglycoprotein-specific Periodic acid-Schiff's (PAS) stain. The new SGPs could be observed only after the partial purification of membrane fractions named as PAS-HI with molecular weight (Mr) 190 kDa and PAS-HII 150 kDa in human, PAS-BIA in buffalo and PAS-PIA and PAS-PIVA in pig. The gels were also stained with Coomassie brilliant blue (CBB) and Silver stain to check the contamination of other membrane proteins in the purified fractions. The quantitative distribution of SGPs was also determined by densitometry. Present study indicates that there are some basic differences in mammalian erythrocyte membrane SGPs, especially with respect to their number and molecular weights indicating major structural variations.

The Hydrolysis of Dimethyl-cis-1,3-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylate by Immobilized Whole Cells of Chromobacterium chocolatum (고정화된 Chromobacterium chocolatum의 Whole Cell을 이용한 Dimethyl-cis- 1,3-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylate의 가수분해)

  • Lee, Youn Jin;Shim, Sang Kyun;Ahn, Yong Hyun
    • Journal of the Korean Chemical Society
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    • v.41 no.9
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    • pp.483-487
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    • 1997
  • The whole cells of Chromobacterium chocolatum was immobilized in the matrix of polyacrylamide and then used for the hydrolysis of dimethyl-cis-1,3-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylate. This hydrolysis yielded the optically active monoester ( > 96% ee) which is useful as an synthetic intermediate of (+)-biotin. We have studied the optimum condition of hydrolysis by using immobilized cells under variable concentration of substrate, reaction time and pH levels. The activity of lipase in immobilized cell was retained for longer than 4 weeks. The best conversion yield of product was obtained when 2 g of wet cell was immobilized and then reacted with 200 mg of substrate at pH 7.

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Cloning and Expression of the Structural Gene for Alcohol Dehydrogenase of Zymomonas mobilis in Escherichia coli (Zymomonas mobilis 알코올 탈수소 효소 유전자의 Cloning과 Escherichia coli 에서의 발현)

  • Yoon, Ki-Hong;Shin, Byung-Sik;M.Y Pack
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.301-306
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    • 1989
  • A genomic library of Zymomonas mobilis DNA was constructed in Escherichia coli using plasmid pUC9 Allyl alcohol was used to screen a genomic clone expressing alcohol dehydrogenase. The plasmids isolated from two clones, which were sensitive to allyl alcohol, were found to be related and to share a common 2.6 kb fragment encoding alcohol dehydrogenase II identified as one of two isozymes in Z. mobilis by staining for alcohol dehydrogenase activity on polyacrylamide gel and spectrophotometric analysis of several substrate oxidations.

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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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Development of Solid-Gelatin Microcarrier for Large Scale Production of Anchorage-Dependent Animal Gell Lines (부착성 동물세포의 대량배양을 위한 젤라틴 미립담체의 개발)

  • 박정극
    • KSBB Journal
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    • v.4 no.1
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    • pp.18-20
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    • 1989
  • Solid gelatin microcarrier with the size distribution between $100{\mu}{\textrm}{m}$ and $400{\mu}{\textrm}{m}$ was prepared for the attachment and growth experiment for anchorage-dependent animal cell lines, i.e., L 929 and BHK 21. The growth and the maximum cell densities on this gelatin based and polyacrylamide (PAA) microcarriers were compared with those on the commercial dextran based Cytodex 3 microcarrier. Both cell lines showed good comparable attachment and growth patterns on the above three microcarriers. The mouse fibroblast, L 929 showed about the same maximum cell density on PAA, gelatin and Cytodex 3 MC'S BUT BHK 21, the baby hamster kidney cell line, showed the best result on Cytodex 3, which was about $4\times10^6$ cells/ml with the microcarrier concentration of 10 g/1.

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Physiology of Strobilus Initiation in Slash Pine -IV. Peroxidase Activity during the Period of Female Strobilus Initiation (슬래쉬 소나무의 화아원기형성(花芽原基形成)의 생리적(生理的) 연구(硏究) -IV. 자화형성기간(雌花形成期間)동안의 정아의(頂芽) PEROXIDASE의 변화(変化))

  • Lee, Kyung Joon
    • Journal of Korean Society of Forest Science
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    • v.50 no.1
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    • pp.5-9
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    • 1980
  • With polyacrylamide gel electrophoresis, peroxidase isoenzyme patterns in the terminal buds of grafted, 20-year-old Pinus elliottii trees were compared between abundant-flowering and poor-flowering trees during the period of female flower bud initiation. A total of nine peroxidase bands were observed in the terminal buds. The total amount of peroxidase enzymes in slash pine buds showed no significant difference between the abundant-flowering and poor-flowering trees. However, the fifth band from the gel front was observed in all the samples of abundant-flowering trees throughout the sampling period (July 8 to September 17), while this band was absent in most of the poor-flowering trees.

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Purification and Characterization of Vitellin from the Red Flour Beetle, Tribolium castaneum Herbst

  • Kim, Seong-Ryul;Choo, Young-Moo;Lee, Seong-Jin;Jin, Byung-Rae;Kim, Jeong-Ho;Heo, In-Bum;Shon, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.1
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    • pp.55-59
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    • 2001
  • The vitellin of the red flour beetled Tribolium castaneum Herbst was purified and characterized. The vitellin of T. castaneum was purified by the FPLC techniques, anion exchange chromatography and gel permeation chromatography. In native-polyacrylamide gel electrophoresis, vitellin of T. castaneum was detected as a single band. This native vitellin has molecular weight of 440 kDa. The vitellin of T. castaneum is composed of three polypeptides, designated Vnl (178 kDa), Vn2 (168 kDa) and Vn3 (52 kDa) in SDS-polyacrylamide gel electrophoresis. Three subunits of vitellin were presented in the female adult hemolymph and egg extracts, but not observed in the male. These three polypeptides gradually decreased during embryogenesis. Polyclonal antiserum raised against purified vitellin reacted with the three polypeptides, Vnl, Vn2 and Vn3. Antisera raised against Vn1 and Vn2 cross-reacted with the two large subunits, Vnl and Vn2, respectively. Another subunits Vn3, however, was not cross-reacted with these two antisera. Also, antiserum raised against Vn3 did not cross-react with the Vn1 and Vn2.

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Characterization of 65 kD Protein in Latex Excreted from Euphorbia lathyris (Euphorbia lathyris에서 분비되는 Latex 65kD 단백질의 특성규명)

  • Park, Hee-Sung
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.319-323
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    • 2004
  • Soluble latex protein fraction excreted from Euporbia lathyris laticifer was resolved by 10% SDS-polyacrylamide gel electrophoresis to identify distinctively displayed latex major protein bands including ELp65, ELp55, ELp43, ELp32 and ELp23. Among them, ELp65 was purified by ammonium sulfate precipitation, gel permeation chromatography and ion exchange chromatography. Its N-terminal amino acid sequencing revealed its homology to the leading region of mature peptide of tomato p69a subtilisin-like protease, suggesting a certain role involved in plant defense system. In the analysis of Southern blot hybridization using PCR-amplified tomato p69a probe DNA, E. lathyris genome was suggested to have a gene family consisting of 3-5 gene members putatively encoding subtilisin-like proteases.

Comparison of soluble antigens from Leptospira interrogans serovars by SDS-PAGE, Crossed Immunoelectrophoresis and Immunoblotting (SDS-PAGE, Crossed Immunoelectrophoresis 및 Immunoblotting을 이용한 Leptospira interrogans 혈청형간 항원 비교)

  • Baik, Yeong-ok;Mah, Jum-sool
    • Korean Journal of Veterinary Research
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    • v.32 no.2
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    • pp.195-205
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    • 1992
  • The soluble antigen profiles and antigenic specificities of Leptospira interrogans serovars icterhaemorrhagiae, canicola, pomona and hardjo were examined by SDS-polyacrylamide gel electrophoresis, crossed immunoeletrophoresis and immunoblotting. The profiles of protein, glycoprotein and fraction containing N-acetylglucosamine of 4 serovars were compared. The protein profiles of 4 serovars were very similar except the range of 14,400 to 30,000 daltons. Molecular weight of glycoprotein of L, pomona was lower than other serovars. L canicola showed extra N-acetylglucosamine bands having molecular weight of 82,000 and 90,000 daltons. In crossed immunoelectrophoresis, a close antigenic relationship was found between L icterohaemorrhagiae and L canicola. In immunoblottings conducted with soluble antigens and rabbit antisera of 4 serovars, Leptospira interrogans serovars possessed cross-reactive antigens and serovar-specific antigens. The molecular weights of serovar-specific antigens were 45,000, 82,000 and 90,000, 31,000 and 24,000 daltons in L icterohaemorrhagiae, L canicola, L pomona, and L hardjo, respectively.

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Acridine orange-induced changes in the O-side chain lengths of lipopolysaccharides of Salmonella pullorum (Salmonella pullorum lipopolysaccharide의 acridine organe에 의한 O-side chain 길이의 변화)

  • Kim, Jong-bae;Kim, Tae-ue;Lee, Won-yong;Yang, Se-whan
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.87-91
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    • 1993
  • The morphological hetergeneity of lipopolysaccharides(LPSs) and variation in the O-side chain lengths of LPSs of Salmonella pullorum, which was serially subcultured on the brain heart infusion agar containing $100{\mu}g/m{\ell}$ of acridine orange, was analyzed in Sephadex G-50 column chromatography and silver-stained polyacrylamide gels. The biochemical differences in LPS W and LPS A0150 were identified. Increases in the contents of O-polysaccharides of LPS A0150 than those of LPS W were reflected in the profiles of chromatography and silver-stained polyacrylamide gels. In summary, LPS molecules of S pullorum A0150 appeared to be enriched in the molecules with long O-polysaccharide chains than those of LPS W.

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