• Title/Summary/Keyword: 2-Dimensional Electrophoresis

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A Simple Method for Detection of Trypsin Inhibitors in Soybean (Glycine max) (대두 Trypsin Inhibitor의 간이검정법)

  • Jo, Ku-Hyung;Lee, Chun-Yung;Hong, Jong-Uek;Kim, In-Soo
    • Korean Journal of Food Science and Technology
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    • v.18 no.5
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    • pp.339-344
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    • 1986
  • The specific reaction of trypsin inhibitors with trypsin to form stable complexes was successfully applied for detection of trypsin inhibitors in soybean. Soybean extract was treated with $Ca^{++}$ to remove globulin fraction, followed by digestion with trypsin and fractionated by chromatography on Sephadex G-50. The void volume fraction contained the trypsin-trypsin inhibitor complexes as well as trypsin. The trypsin inhibitors were then detected by their molecular weight differences on SDS-polyacrylamide gel electrophoresis, in which the complexes dissociate into trypsin and its inhibitors. With the method proposed, trypsin inhibitors were indentified by their ability forming the stable complexes with trypsin and their anti-tryptic moiety. The formation of the complexes with trypsin was further confirmed by two dimensional electrophoresis and DEAE-Sephadex A-25 chromatography. Employing the proposed method, it was found that soybean (Glycine max cv. Hill) contained 7 trypsin inhibitors.

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Identification of Key beta-1,3-glucan Synthesis Enzymes in Agrobacterium sp. ATCC31750 (Agrobacterium sp. ATCC31750에 대한 beta-l,3-glucan 합성 대사경로의 주요 단백질 검출)

  • Kim Ryo Hwa;Lee Jung Heon
    • KSBB Journal
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    • v.19 no.5
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    • pp.406-409
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    • 2004
  • Matrix Assisted Laser Desorption ionization Time of Flight (MALDI-TOF) was used for enzymes identification related to B -1,3-glucan synthesis. Agrobacterium sp. ATCC31750 was cultivated with two stage Continuous Stirrer Tank Reactor (CSTR) and the cells were harvested and their protein profiles were analysed by two dimensional electrophoresis. The specific enzyme spot was treated with trypsin and ana lysed by MALDI-TOF to get peptide molecular weight. The peptide molecular weights were matched with Agrobacterium tumefacience's Data Base from the matrix science site, then could identify the avaliable key enzymes. In this study, we identified key metabolite of synthesis of beta-1,3-glucan, such as glucose-6-phosphate isomerase, phosphoglucomutase, B-1,3-glucan synthase and glucokinase, and we also identified uracil phosphoribocyl transferase and Ribosome recycling factor also.

Apnea of Somatic Cell Cloned Piglets with Congestion is Caused by Cardiopulmonary

  • Lee, So-Young;Park, Mi-Ryeung;Cho, Seong-Keun;Park, Yun-Jung;Kwon, Deug-Nam;Lee, Eun-Kyeong;Son, Woo-Jin;Kim, Jin-Hoi
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.186-186
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    • 2004
  • In this study, we generated 40 somatic cell cloned (scNT) piglets. Of these, three displayed congestion in both liver and lung, and died within the first week of life. Two-dimensional gel electrophoresis experiments revealed changes in the responses of several detoxification-related proteins to stress and inflammation. As a result, congestive livers and lungs displayed extensive hepatopneumonic apoptosis.(omitted)

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A Proteomic Approach to Study msDNA Function in Escherichia coli

  • Jeong, Mi-Ae;Lim, Dongbin
    • Journal of Microbiology
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    • v.42 no.3
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    • pp.200-204
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    • 2004
  • Retron is a prokaryotic genetic element that produces multicopy single-stranded DNA covalently linked to RNA (msDNA) by a reverse transcriptase. It was found that cells producing a large amount of msDNA, rather than those that did not, showed a higher rate of mutation. In order to understand the molecular mechanism connecting msDNA production to the high mutation rate the protein patterns were compared by two dimensional gel electrophoresis. Ten proteins were found to be differentially expressed at levels more than three fold greater in cells with than without msDNA, nine of which were identified by MALDI TOF MS. Eight of the nine identified proteins were repressed in msDNA-producing cells and, surprisingly, most were proteins functioning in the dissimilation of various carbon sources. One protein was induced four fold greater in the msDNA producing cells and was identified as a 30S ribosomal protein S2 involved in the regulation of translation. The molecular mechanism underlying the elevated mutation in msDNA-producing cell still remains elusive.

IDENTIFICATION OF DIFFERENTIALLY EXPRESSED PROTEINS IN DIFFERENT GROWING STAGES IN CHICKEN LIVER BY PROTEOMICS APPROACH

  • Lee, K.Y.;Jung, K.C.;Jang, B.G.;Choi, K.D.;Lee, J.H.
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2006.11a
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    • pp.74-76
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    • 2006
  • 닭의 간은 해독작용, 당의 저장, 혈장 단백질의 합성 등 주요 기능을 하는 것으로 알려져 있다. 본 연구는 간에서 성장 단계별로 발현량에 차이를 보이는 단백질들을 비교해 보았다. 2차원적 전기 영동에 의해 분리된 300개 이상의 단백질들이 확인되었으며 이 중 성장 단계별 특이적인 13개의 단백질은 MALDI-TOF MS에 의하여 분석이 되어졌다. 본 연구를 통하여 밝혀진 단백질들은 생화학적인 연구에 중요한 자료를 제공할 것으로 사료된다.

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Identification and Partial Purification of Two Hydrogenase Isoenzymes from Escherichia coli (대장균으로부터 두 종류의 수소발생 동위효소의 확인과 부분정제)

  • 최석정;양철학
    • Korean Journal of Microbiology
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    • v.29 no.5
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    • pp.296-300
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    • 1991
  • The membrane-bound Escherichia coli hydrogenases were purified partially by the solubilization with detergents. the E. coli crude extract was solubilized with sodium deoxycholate and dialyzed against the buffer containing Triton X-100. Two different hydrogenases were obtained by the DEAE-cellulose, hydroxyapatite and Sephedex G-200 column chromatography. The one was unstable during purification and contained 70- and 47-kDa polypeptides as major proteins. The other showed high H2-evolving activity and had major polypeptides of Mr 31 and 27. Those polypeptides were detected by the two-dimensional electrophoresis.

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Proteomic Analysis of Drought Stress-Responsive Proteins in Rice Endosperm Affecting Grain Quality

  • Mushtaq, Roohi;Katiyar, Sanjay;Bennett, John
    • Journal of Crop Science and Biotechnology
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    • v.11 no.4
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    • pp.227-232
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    • 2008
  • Drought stress is one of the major abiotic stresses in agriculture worldwide. We report here a proteomic approach to investigate the impact of post-fertilization drought on grain quality in rice seed endosperm (Oryza sativa cv. IR-64). Plants were stressed for 4 days at 3 days before heading. Total proteins of endosperm were extracted and separated by two-dimensional gel electrophoresis. Not many protein spots showed differential accumulation in drought-stressed samples. More than 400 protein spots were reproducibly detected, including three that were up-regulated and five down-regulated. Mass spectrometry analysis and database searching helped us to identify six spots representing different proteins. Functionally, the identified proteins were related to protein synthesis and carbohydrate metabolism, such as Granule-Bound Starch Synthase (GBSS, Wx protein), which is thought to play a very important role in starch biosynthesis and quality, a very crucial factor in determining rice grain quality.

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Microbial Diversity Information Facility: Bacteriology Insight Orienting System (BIOS)

  • Shimura, Junko;Shimiz, Hideyukiu;Tsuruwaka, Keiji;Moritani, Yukimitsu;Miyazaki, Kenji;Tsugita, Akira;Watanabe, Makoto M.
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.135-141
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    • 2000
  • Global Biodiversity is common interest of humans for better health and sustainable development of the society. To provide access and analysis on microbial diversity information, Bacteriology Insight Orienting System (BIOS) has been developed. BIOS contains 6402 species and subspecies names of bacteria and archaea, 2606 names of cyanobacteria by March 2000. BIOS of which web based analytical tool provides windows to compare the results of phylogenetic analysis based on 16S rDNA sequence and the results of cluster analysis on proteome profiling. The sequence data and 2 dimensional gel electrophoresis analysis data were accumulated in BIOS database content for cyanobacteria reclassification and taxonomy. (BIOS URL: http.://www-sp2000ao.nies.go.jp/bios/index.html).

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