• Title/Summary/Keyword: Isoelectric focusing

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Electrophoretical Analysis of 36-Kilodalton Outer Membrane Protein of Vibrio vulnificus ATCC 27562

  • Moon-Soo Heo;Cho-Rok Jung
    • Journal of Life Science
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    • v.9 no.1
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    • pp.35-39
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    • 1999
  • Elecrophoreticl analysis of a 36 kDa protein was runned by SDS-PAGE, isoelectric focusing (IEF) and two dimensional electrophoresis pattern. Major 36 kDa and 25, 46, 48, 66 kDa protein were detected by Coomassie blue stain on SDS-PAGE. Major 36kDa protein was eluted for production of antiserum for serological analysis, IEF and two dimensional electrophoresis. Isoelectric point of 36kDa was aout pH 8.5. Two dimensional electrophoresis of eluted 36kDa showed one point on the gel. Anti-36 kDa serum made by newzilland rabbit for serological test. In ELISA, final titer of antibody was 100×{TEX}$2^5}${/TEX} : 1. Neutralize ability of serum was examined by slide agglutination test and colonization test in rat. Anti-36 kDa serum agglutinated whole cell of V. vulnificus were inhibited colonization on intestine in rat. Accordingly In this paper contain some electrophoretical analysis and serological test of a 36 kDa OMP of V. vulnificus.

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Development of a Four-way Interface for Online Capillary Isoelectric Focusing-Electrospray-Mass Spectrometry (CIEF-ESI-MS)

  • Yu, Hai Dong;Kim, Byungjoo;Shin, Dae-Ho;Ahn, Seonghee
    • Mass Spectrometry Letters
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    • v.4 no.4
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    • pp.83-86
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    • 2013
  • A new interface for coupling CIEF and MS using a four-way cross has been developed in a single mechanical system. This new interface could be operated without the electric discontinuity and reinstallation of lines. Additionally, a bare fused silica capillary was facilitated as a spray needle to produce electrospray and to guide catholyte or sheath liquid. Focusing for CIEF was completed in a hanging droplet at the end of spray needle. This capillary spray needle also provided stable spray, enhanced the ionization efficiency and increased sensitivity. Results with carbonic anhydrase I showed that focusing and spraying were well completed with the new interface and the new spray needle.

Purification and Characterization of Arginase from Schizosaccharomyces pombe

  • Kang, Jung-Hoon
    • BMB Reports
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    • v.28 no.3
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    • pp.232-237
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    • 1995
  • Arginase was purified to homogeneity from Schizosaccharomyces pombe. The purified enzyme is a tetramer with a subunit molecular weight of 42,000. Activity is optimal at pH 10.0 and at $60^{\circ}C$ The enzyme migrated during isoelectric focusing showing a pl=5.4. The enzyme exhibited hyperbolic kinetics at pH 10.0 with an apparent $K_m$ for L-arginine of 18 mM. Arginase activity was strongly inhibited by L-glutamate.

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Purification and Characterization of ATPase and Phosphatase of Light Membrane Vesicles Isolated from Cucurbita pepo (Cucurbita pepo에서 분리한 Light Membrane Vesicle의 ATPase와 Phosphatase의 정제 및 특성)

  • 오승은
    • Journal of Plant Biology
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    • v.33 no.4
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    • pp.325-332
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    • 1990
  • Light membrane vesicles were isolated from the zucchini hypocotyl by floatation on ficoll density gradients and the proteins were solubilized with Triton X100. Three ATP-hydrolyzing enzymes were partially purified by ion-exchange and gel filtration chromatography and isoelectric focusing. There are plasma membrane-type ATPase whose activity was inhibited by vanadate but not by nitrate, tonoplast-type ATPase which was sensitive to nitrate but insensitive to vanadate and one having a phosphatase activity with a pI value different from that of an acid phosphatase. A fraction was obtained after DEAE-ion-exchange chromatography crossreacting with polyclonal antibodies against Ca2+ -ATPase from human erythrocytes.

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잠란의 휴면특이 peptides 검출을 위한 2D-전기 영동 분석

  • 고현정;김현수;김영태;강석우;황재삼;성수일
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.04a
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    • pp.43-43
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    • 2003
  • 잠란의 휴면 개시, 유지 및 각성 등에 따른 단백질의 변화를 알아보기 위해 2D-전기영동에 의한 peptides 분석을 하였다. 잠품종은 백옥잠을 사용하였으며, 단백질 분석은 산란 후 5일 경과란(휴면란), 산란 후 20시간에 침산하여 1일 및 2일 경과란, 및 산란 후 2일 경과 후 냉장(5$^{\circ}C$)처리하여 1일, 3일, 5일된 잠란을 각각 대상으로 하였다. 2D-전기영동은 1차로 pH3-10 range에서 isoelectric focusing 하고 2차로 SDS-PAGE한 후 silve stainin으로 peptides를 검출하였다. (중략)

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Studies on Agronomic and Biochemical Characteristics in Korean Native Rice Cultivars I. Isozyme Variation by Isoelectric Focusing - EST, ME, PGI, HK (한국 벼 재래종의 작물학적 및 생화학적 특성연구 I. 등전점 전기영동법에 의한 Esterase, Malic enzyme, Phosphoglucose isomerase 및 Hexokinase 동위효소 형태 특성)

  • Eun M. Y.;Kim Y. K.;Cho Y. G.;Kim Y. W.;Chung T. Y.;Choi H. C.
    • Proceedings of the Korean Society of Crop Science Conference
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    • 1989.05a
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    • pp.18-19
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    • 1989
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Comparative Studies of Invertase Isozymes Produced by Rhodotorula glutinis K-24 (Rhodotorula glutinis K-24가 생산하는 Invertase Isozymes군에 관한 비교 연구)

  • Lee, Tae-Ho;Kim, Chul;Lee, Sang-Ok
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.313-320
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    • 1989
  • Rhodotorula glutinis K-24 was found to produce internal, cell wall bound, and external invertase. Internal invertase was purified by column chromatographies on DEAE-Sephadex A-50, Sp-sephadex C-50, gel filtration on Sephadex G-200 and isoelectric focusing. Cell wall bound invertase was partially purified by the following procedures; column chromatography on DEAE-Sephadex A-50 and gel filtration on Sephadex G-100. Optimum pH and temperature for enzymatic activities of internal and cell wall bound invertase were pH 3.0 and 6$0^{\circ}C$, respectively. Both enzymes were inhibited by HgC1$_2$, AgNO$_3$, MnSO$_4$, and sodium dodecylsulfate. The molecular weights of internal and cell wall bound invertases were estimated to be 310,000 and 61,000, respectively. Other physicochemical properties of the both enzymes were similar.

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Genetic Polymorphism of Plasma Vitamin D-Binding Protein (Gc) in Some Asian Sheep

  • Tsunoda, K.;Doge, K.;Hasnath, M.A.;Rajbhandary, H.B.;Xu, W.;Zhanchiv, T.;Chau, B.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.3
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    • pp.318-322
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    • 1998
  • Using polyacrylamide-gel isoelectric focusing followed by immunoblotting, genetic polymorphism of plasma vitamin D-binding protein (Gc) was examined in Asian sheep. The Gc polymorphism was revealed in the Khalkhas sheep of Mongolia, consisting of F, S and W variants, and the Yunnan native sheep of China, consisting of F and S variants. In particular, W was a new variant. The V variant detected in European sheep up to now was not observed in these sheep. The Bhyanglung, Baruwal, Kagi and Lampuchhre sheep of Nepal and local sheep of Bangladesh and Vietnam were monomorphic for the S variant. Family data and population genetic data supported the hypothesis that these variants were controlled by codominant alleles. In these Asian sheep, distribution of the $Gc^s$ allele was predominant (0.9571-1) and was seen as well in European sheep (Suffolk, Corriedale, Cheviot and Finnish Landrace) raised in Japan. $Gc^w$ allele was detected only in the Khalkhas sheep with the low frequency of 0.0025. The $Gc^v$ allele was detected in the Suffolk and Corriedale sheep (0.0080 and 0.0682), but not in any of the Asian sheep studied.

Isoenzyme patterns and phylogenetic relationships in Acanthamoeba spp. isolated from contact lens containers in Korea

  • Shin, Ho-Joon;Cho, Myung-Soo;Kim, Han-jip;IM, Kyung-Il
    • Parasites, Hosts and Diseases
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    • v.37 no.4
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    • pp.229-236
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    • 1999
  • In order to refer to the basic information regarding the identification of isolates obtained from a contact lens container in Korea, the isoelectric focusing gel electrophoresis was employed to compare the isoenzyme band patterns among Acanthamoeba spp. including eight isolates and the simple pairwise dissimilarity analysis was carried out. For an alkaline phosphate development, isolate 7 and Acanthamoeba polyphaga showed homologous band patterns, and isolates 1, 2, and 3 showed the same patterns. For lactate dehydrogenase, similar patterns were observed in isolates 2 and 3. Isolates 3 and 5 showed homologous band patterns for malate dehydrogenase and glucose phosphate isomerase. For hexokinase, isolates 4, 7, and A. hatchetti showed the same band patterns. In others, a considerable number of interstrain polymorphisms was observed in nine isoenzyme band patterns. In Acanthamoeba group II, genetic distances among isolates 1, 2, 3, 4, and 5 ranged from 0.104 to 0.200. In comparison to A. castellanii, A. hatchetti, and A. poIyphaga, genetic distances of isolates 7 and 8 were 0.254 and 0.219, respectively. In Acanthamoeba group III, including A. culbertsoni, A. healyi, and A. royreba, isolate 6 had genetic distances which ranged from 0.314 to 0.336. Finally, when comparing to the six reference Acanthamoeba, it was possible to classify isolates 1, 2, 3, 4, and 5, as genetically close-related species and as independent species group. Furthermore, isolates 6, 7 and 8 were identified as independent species as well.

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