• 제목/요약/키워드: isoelectric focusing

검색결과 129건 처리시간 0.022초

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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    • 제9권1호
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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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    • 제4권4호
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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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    • 제28권3호
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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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Cucurbita pepo에서 분리한 Light Membrane Vesicle의 ATPase와 Phosphatase의 정제 및 특성 (Purification and Characterization of ATPase and Phosphatase of Light Membrane Vesicles Isolated from Cucurbita pepo)

  • 오승은
    • Journal of Plant Biology
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    • 제33권4호
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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
    • 한국잠사학회:학술대회논문집
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    • 한국잠사학회 2003년도 제46회 춘계 학술연구 발표회
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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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한국 벼 재래종의 작물학적 및 생화학적 특성연구 I. 등전점 전기영동법에 의한 Esterase, Malic enzyme, Phosphoglucose isomerase 및 Hexokinase 동위효소 형태 특성 (Studies on Agronomic and Biochemical Characteristics in Korean Native Rice Cultivars I. Isozyme Variation by Isoelectric Focusing - EST, ME, PGI, HK)

  • 은무영;김용권;조용구;김영우;정태영;최해춘
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 1989년도 하계 학술대회지
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    • pp.18-19
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    • 1989
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Rhodotorula glutinis K-24가 생산하는 Invertase Isozymes군에 관한 비교 연구 (Comparative Studies of Invertase Isozymes Produced by Rhodotorula glutinis K-24)

  • Lee, Tae-Ho;Kim, Chul;Lee, Sang-Ok
    • 한국미생물·생명공학회지
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    • 제17권4호
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    • pp.313-320
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    • 1989
  • 세포내와 세포벽에 invertase가 존재하며 catabolite inhibition을 받지않고 세포외로 구성적으로 invertase를 생산하는 Rh. glutinis K-24를 공시균주로 선택하여 세포내와 세포벽 invertase를 정제한 후 정제효소의 효소화학적 성질을 밝혔다. 세포내 invertase는 DEAE-Sephadex A-50에 의한 Batchment, DEAE-Sephadex A-50 column chromatography, gel filtration on Sephadex G-200, isoelectric focusing 등의 조작에 의해 Disc 전기영동상 단일한 효소단백질까지 정제되었다. 세포벽 invertase는 B. pumilis에 의해 생산되는 세포벽 용해효소를 작용시켜 invertase를 용출 시킨 다음 DEAE-Sephadex A-50 column chromatography, gel filtration on Sephadex G-100의 조작에 의해 부분 정제하였다. 세포내와 세포벽 Invertase의 분자량은 각각 약 310,000과 67,000이었고 세포내 경우는 subunit 분자량이 약 70,000이었다. 세포내와 세포벽 효소의 반응 최적 pH와 온도는 모두 4.0와 6$0^{\circ}C$였으며 온도 안정성도 7$0^{\circ}C$까지 거의 비슷하게 나타났다. 그러나 Km값은 세포내 효소의 경우는 4.3x$10^{-3}$M이었고 세포벽 효소의 경우는 2.1x$10^{-2}$M이었다. 그외 Inhibitor에 대한 효소의 저해양식도 양효소에 비슷한 결과를 나타내었다.

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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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    • 제11권3호
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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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    • 제37권4호
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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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