• Title/Summary/Keyword: Two-Dimensional Gel Electrophoresis

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Differential Induction of Pathogenesis-Related Proteins in the Compatible and Incompatible Interactions of Tomato Leaves with Xanthomonas campestris pv. vesicatoria (Xanthomonas campestris pv. vesicatoria와 토마토잎의 친화적, 불친화적 반응에서 병생성관련 단백질의 유도)

  • 김정동;황병국
    • Korean Journal Plant Pathology
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    • v.11 no.1
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    • pp.53-60
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    • 1995
  • Inoculation with the compatible strain Ds 1 of Xanthomonas campestris pv. vesicatoria caused brownish ad water-soaked lesions, but incompatible strain Bv5-4a produced hypersensitive symptoms with local necrosis on tomato (cv. Kwangyang) leaves. Bacterial populations of the compatible strains Ds 1 propagated more greatly than the incompatible strain Bv5-4a at the frist onset, but no differences were observed 5 days after inoculation. The bacterial infection induced the synthesis and accumulation of soluble proteins in tomato leaves, especially in the incompatible interaction. Native-polyacrylamide gel electrophoresis distinguished the soluble proteins in the tomato leaves infected by the compatible or incompatible strains. A protein of low molecular weight occurred only in the incompatible interaction. Some pathogenesis-related (PR) proteins, especially the 15, 18, 23, 26 and 54 kDa proteins, were detected only in the infected tomato leaves. In the two-dimensional electrophoresis, some proteins with different molecular weights (Mr. 21∼29 kDa) and the pI 8∼9 appeared more distinctly only in the incompatible interaction. These data suggest that the de novo synthesis of some PR proteins in tomato may be significant in defense against X. c. pv. vesicatoria.

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Identification of the Proteins Expressed at Optimum Marbling in Hanwoo Loin (한우육의 육질에 중요 영향을 미치는 근내지방축적 최적기에 발현되는 단백질 동정)

  • 임진규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.3
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    • pp.579-585
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    • 1999
  • Proteins from the loin tissues age ranged from 0 to 24 months of ten Korean cattle were extracted, separated and compared on two dimensional(2 D) gels to identify the proteins whose expression is highly correlated to marbling. We also compared the difference of loin proteins between castrated and non castrated bull cows on two dimensional gels. As the marbling in the loin of the cattle is optimized at 18 to 24 months, eight proteins expressed significantly higher level in 24 month than in 0 or 6 month were selected in terms of isoelectric points(pIs) and molecular weights. Using these values, we searched the Swiss Prot database via the ExPASy molecular biology server with TagIdent program. The proteins with the nearest molecular weights and isoelectric points were selected from the lists. These possible candidates were confirmed by N terminal microsequencing of the eight selected proteins. Three proteins, myoglobin, hemoglobin and ATPase, whose N termini were not blocked could be microsequenced and found to be exactly matched to the selected candidates. It is suggested that the proteins increasingly expressed in marbling periods can be involved in meat color, lipid transport and flavor improvement.

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Two-Dimensional Reference Map of Schizosaccharomyces pombe Proteins (Update)

  • Kim, Sun-Kyung;Won, Mi-Sun;Sun, Nam-Kyu;Jang, Jae-Won;Lee, Seung-Hee;Shin, Hee-Young;Song, Kyung-Bin
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1499-1512
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    • 2006
  • Based on the first 2D reference map of the fission yeast Schizosaccharomyces pombe protein reported previously, we expanded and updated the map using narrower pI ranges. In this paper, 240 protein spots were identified on our reference map. In the pI 4-7 range, 144 spots corresponding to 86 different proteins were identified. In the pI 6-9 range, 43 spots corresponding to 35 different proteins were identified. Fifty-three new spots corresponding to 39 different proteins were further identified in the pI 5-6 range.

Introduction of Ti Plasmid into Bradyrhizobium japonicum by Spheroplast Transformation (형질전환(形質轉換)에 의한 Ti Plasmid의 Bradyrhizobium japonicum에의 도입(導入))

  • Yun, Han-Dae;Cho, Moo-Je;Park, Chan-Young;Lee, Ke-Ho
    • Applied Biological Chemistry
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    • v.30 no.4
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    • pp.293-299
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    • 1987
  • Bradyrhizobium japonicum spheroplasts were prepared by culturing cells in the presence of glycine, follwed by treatment with lysozyme. The cells were examined by electron microscopy during the formation of spheroplast. Then Ti plasmid from Agrobacterium tumefaciens 15955 was introduced into Bradyrhizobium japonicum by glycine-lysozyme induced spheroplast transformation. After cell wall regeneration, transformants were selected by the ability of utilization of octopine. Transformation were received at a frequency of $1{\times}10^{-7}$. The transformants obtained from spheroplast transformation harbored the introduced Ti plasmid, which was identified by agarose gel electrophoresis. Furthermore, the differences in their gene products were observed between the transformant and the recipient cell by two-dimensional polyacrylamide gel electrophoresis. The transformants which still possessed the same ability nodulate soybean (Glycine max.) as that of the original host strain, acquired the ability to induce tumor on Petunia hybrida like Agrobacterium, but formed the small crown galls in size compared to those of Agrobacterium tumefaciens.

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Separation and Characterization of Spikelet Proteins at Young Microspore Stage in Rice

  • Woo, Sun-Hee;Kim, Tae-Seon;Cho, Seong-Woo;Ahn, Jung-Gu;Chung, Keun-Yook;Lee, Byung-Moo;Cho, Yong-Gu;Kim, Hong-Sig;Song, Beom-Heon;Lee, Chul-Won;Jong, Seung-Keun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.1
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    • pp.66-72
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    • 2006
  • Spikelet proteins expressed at the young microspore stage in rice were separated and analysed by two-dimensional polyacrylamide gel electrophoresis (2DE). The separated proteins were electro blotted onto a polyvinylidene difluoride (PVDF) membrane, and 50 proteins were analyzed by a gas-phase protein sequencer. The N-terminal amino acid sequences of 20 out of 50 proteins were determined. N-terminal regions of the remaining proteins could not be sequenced because of blocking. The internal amino acid sequences of proteins were determined by sequence analysis of peptides obtained by the Cleveland peptide mapping method. Results revealed the presence of the photosynthetic apparatus at rice young microspore stage. Major proteins identified in this study could be used as a marker for various studies on physiological stresses.

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.

Proteomic Studies in Plants

  • Park, Ohk-Mae K.
    • BMB Reports
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    • v.37 no.1
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    • pp.133-138
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    • 2004
  • Proteomics is a leading technology for the high-throughput analysis of proteins on a genome-wide scale. With the completion of genome sequencing projects and the development of analytical methods for protein characterization, proteomics has become a major field of functional genomics. The initial objective of proteomics was the large-scale identification of all protein species in a cell or tissue. The applications are currently being extended to analyze various functional aspects of proteins such as post-translational modifications, protein-protein interactions, activities and structures. Whereas the proteomics research is quite advanced in animals and yeast as well as Escherichia coli, plant proteomics is only at the initial phase. Major studies of plant proteomics have been reported on subcellular proteomes and protein complexes (e.g. proteins in the plasma membranes, chloroplasts, mitochondria and nuclei). Here several plant proteomics studies will be presented, followed by a recent work using multidimensional protein identification technology (MudPIT).