• Title/Summary/Keyword: two-dimensional gel

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Proteomic analysis of rice mutants susceptible to Magnaporthe oryzae

  • Ryu, Hak-Seung;Song, Min-Young;Kim, Chi-Yeol;Han, Muho;Lee, Sang-Kyu;Ryoo, Nayeon;Cho, Jung-Il;Hahn, Tae-Ryong;Jeon, Jong-Seong
    • Plant Biotechnology Reports
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    • v.3 no.2
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    • pp.167-174
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    • 2009
  • To identify genes involved in rice Pi5-mediated disease resistance to Magnaporthe oryzae, we compared the proteomes of the RIL260 rice strain carrying the Pi5 resistance gene with its susceptible mutants M5465 and M7023. Proteins were extracted from the leaf tissues of both RIL260 and the mutant lines at 0, 24, and 48 h after M. oryzae inoculation and separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis identified eight proteins that were differently expressed between the resistant and susceptible plants (three down- and five up-regulated proteins in the mutants). The down-regulated proteins included a triosephosphate isomerase (spot no. 2210), a 2,3-bisphosphoglycerate-independent phosphoglycerate mutase (no. 3611), and an unknown protein (no. 4505). In addition, the five up-regulated proteins in the mutants were predicted to be a fructokinase I (no. 313), a glutathione S-transferase (no. 2310), an atpB of chloroplast ATP synthase (no. 3616), an aminopeptidase N (no. 3724), and an unknown protein (no. 308). These results suggest that proteomic analysis of rice susceptible mutants is a useful method for identifying novel proteins involved in resistance to the M. oryzae pathogen.

Effects of Colchicine and EMS Dropping to Apical Meristem on Induced Mutation and Plant Protein Pattern in Wasabia japonica Matsum (Colchicine과 EMS 돌연변이 처리가 고추냉이 식물체의 단백질 Pattern에 미치는 영향)

  • Park, Kie-In;Eun, Jong-Seon;Choi, Sun-Yong;Kwon, Sung-Whan;Lee, Kang-Soo;So, Sang-Sup;Youn, Myung-Ja
    • Korean Journal of Medicinal Crop Science
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    • v.5 no.1
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    • pp.36-42
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    • 1997
  • This study was investigated on the effects of colchicine and EMS. induced mutation, plant on the germination and polypeptide pattern and peroxidase activity monitored by two dimensional gel analysis in Wasabia japonica Matsum (wasabi). Germination rate of Muju was higher than that of Ulrung-Do and optimum concentration for germination was appeared 100 ppm GA3 containing with 10 ppm BAP in these cultivars. Survived plants rate of Muju was higher than that of Ulrung-Do after colchicine and EMS treatment. Peroxidase activity and plant height were decreased by mutagen treatments, while incresed in root and stem thickness. The number of protein spots and pattern showed difference between Muju and Ulrung-Do . The plants treated mutagen increased polypeptide spots, especially EMS treatment showed more different polypeptide pattern compared to control.

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Proteomic Analysis of Protein Expression in Streptococcus pneumoniae in Response to Temperature Shift

  • Lee Myoung-Ro;Bae Song-Mee;Kim Tong-Soo;Lee Kwang-Jun
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.375-382
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    • 2006
  • From its initial colonization to causation of disease, Streptococcus pneumoniae has evolved strategies to cope with a number of stressful in vivo environmental conditions. In order to analyze a global view of this organism's response to heat shock, we established a 2-D electrophoresis proteome map of the S. pneumoniae D39 soluble proteins under in vitro culture conditions and performed the comparative proteome analysis to a 37 to $42^{\circ}C$ temperature up-shift in S. pneumoniae. When the temperature of an exponentially growing S. pneumoniae D39 culture was raised to $42^{\circ}C$, the expression level of 25 proteins showed changes when compared to the control. Among these 25 proteins, 12 were identified by MALDI-TOF and LC-coupled ESI MS/MS. The identified proteins were shown to be involved in the general stress response, energy metabolism, nucleotide biosynthesis pathways, and purine metabolism. These results provide clues for understanding the mechanism of adaptation to heat shock by S. pneumoniae and may facilitate the assessment of a possible role for these proteins in the physiology and pathogenesis of this pathogen.

Proteomic Analysis of Rat PC12 Cells Exposed to Cyclosporin A

  • Jung, Ji-Yeon;Seol, Kwang;Jeong, Yeon-Jin;Kim, Won-Jae;Oh, Sang-Jin
    • International Journal of Oral Biology
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    • v.34 no.1
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    • pp.29-36
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    • 2009
  • Cyclosporin A (CsA) has been used clinically as an immunosuppressive drug to prevent organ transplant rejection and in basic research as a mitochondrial permeability blocker. It has been reported that CsA has a protective role in severed neurons and a neurotrophic effect in neuronal cells. However, the molecular mechanisms underlying the stimulation of neuronal cell proliferation by CsA have not yet been elucidated. In our current study, we investigated CsA responsive proteins in PC12 cells using a systematic proteomic approach. The viability of these cells following CsA treatment increased in a dose- and time-dependent manner. Proteins in the CsA-treated PC12 cells were profiled by two-dimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption ionization time-of flight (MALDI-TOF) and electrospray ionization quadupole time-of-flight mass spectrometries (EIQ-TOFMS). This differential expression analysis showed significant changes for 10 proteins (6 up-regulated and 4 down-regulated) upon CsA treatment that were related to cell proliferation, metabolism and the stress response. These proteomics data further our understanding of the proliferation mechanisms of PC12 cells exposed to CsA and demonstrate that our methodology has potential to further elucidate the mechanisms and pathways involved.

A proteomic approach reveals the differential protein expression in Drosophila melanogaster treated with red ginseng extract (Panax ginseng)

  • Liu, Qing-Xiu;Zhang, Wei;Wang, Jia;Hou, Wei;Wang, Ying-Ping
    • Journal of Ginseng Research
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    • v.42 no.3
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    • pp.343-351
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    • 2018
  • Background: Red ginseng is a popularly used traditional medicine with antiaging effects in Asian countries. The present study aimed to explore the changes in protein expression underlying the mechanisms of life span extension and antiaging caused by red ginseng extract (RGE) in Drosophila melanogaster. Methods: A proteomic approach of two-dimensional polyacrylamide gel electrophoresis (2-DE) was used to identify the differential abundance of possible target proteins of RGE in D. melanogaster. The reliability of the 2-DE results was confirmed via Western blotting to measure the expression levels of selected proteins. Proteins altered at the expression level after RGE treatment (1 mg/mL) were identified by matrix-assisted laser desorption/ionization-time of flight tandem mass spectrometry and by searching against the National Center for Biotechnology nonredundant and Uniprot protein databases. The differentially expressed proteins were analyzed using bioinformatics methods. Results: The average survival life span of D. melanogaster was significantly extended by 12.60% with RGE treatment (1 mg/mL) compared to untreated flies. This followed increased superoxide dismutase level and decreased methane dicarboxylic aldehyde content. Based on the searching strategy, 23 differentially expressed proteins were identified (16 up-regulated and 7 down-regulated) in the RGE-treated D. melanogaster. Transduction pathways were identified using the Kyoto Encyclopedia of Genes and Genomes database, and included the hippo and oxidative phosphorylation pathways that play important roles in life span extension and antiaging process of D. melanogaster. Conclusion: Treatment with RGE in D. melanogaster demonstrated that mechanisms of life span extension and antiaging are regulated by multiple factors and complicated signal pathways.

Effects of $Ca^2+$ and Protein Kinase C on the Chick Myoblast Differentiation (Ca$^2+$ 및 Protein Kinase C가 배양한 계배근원세포의 분화에 미치는 영향)

  • 정기화;김세재;박정원;박영철;이정주
    • The Korean Journal of Zoology
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    • v.32 no.1
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    • pp.40-47
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    • 1989
  • Alteration of intracellular calcium ion Concentration by adding of either calcium ionophore A23187 or EGTA in culture medium at 24 hr after cell plating resulted in remarkable changes in the progression of differentiation of chick embryo myoblast. When separated myoblast proteins using two-dimensional gel electrophoresis, synthesis patterns of several proteins changed upon the addition of either A23187 or EGTA. Treatment of A23187 and calciumactivated neutral protease at 24 hr after initial plating caused an increase in the rate of fusion compared to control culture. However, EGTA inhibited the myoblast fusion to a marked degree. A23187 treated at 24hr also increased the activity of protein kinase C during the fusionprogressed period. It seems that intracellular calcium ion plays an important role in the myoblast differentiation in vitro together with the protein kinase C and calcium-activated neutral protease.

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In Vitro Flowering System (In Vitro 시스템에 의한 화호형성)

  • 류장렬;이행순;이광웅
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.213-237
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    • 1987
  • In vitro flowering system may minimize the confounded influence of non-floral meristem parts of plants in studying the relationship of a given treatment and flowering responses. We have induced flower buds from plantlets regenerated from zygotic embryo-derived somatic embryos of ginseng, which circumvented the normal 2-year juvenile period before flowering. The result suggests that the adulthood of ginseng root explants in the experiment previously conducted by Chang and Hsing (1980; Nature 284: 341-342) is not prerequired to flowering of plantlets regenerated through somatic embryogenesis. We have also induced flower buds from elongated axillary brandches from cotyledonary nodes by culturing ginseng zygotic embryos, seedlings, and excised cotyledonary nodes. It was found that 6-benzyladenine (BA) supplemented to the medium was essential for flowering, whereas abscisic acid (ABA) was inhibitory. Gibberellic acid(GA3) was also required for flowering when ABA was present with BA in the medium. The results suggest that cytokinins, gibberellins, and inhibitors play primary, permissive, and preventive roles, respective-ly, in the induction of flowering of ginseng. Tran Thanh Van (1980; Int. Rev. Cytol., Suppl. IIA: 175-194) has developed the "thin cell layer system" in which the induction of shoots, roots, or flower buds from epidermal layer explants were controlled by culture conditions and exogenous growth regulators in the medium, Utilizing the thin cell layer system, Meeks-Wagner et al. (1989; The Plant Cell 1: 25-35) have cloned genes specifically expressed during floral evocation. However, the system is too tedious for obtaining a sufficient amount of plant materials for biochmical and molecular biological studies of flowering. We have developed a garlic callus culture system and one obvious advantaging over the thin cell layer system is that an abundant cells committed to develope into flower buds proliferate. When the above cells were compared by two-dimensional gel electrophoresis with those which have just lost the competence for developing into flower buds, a few putative proteins specific to floral evocation were detected. The garlic callus culture system can be further explored for elucidation of the molecular biological mechanism of floral evocation and morphogenesis.hogenesis.

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Loss of a Strain-Specific Protein by Bacterial Infection in Amoeba proteus (Amoeba proteus에 있어서 박테리아 감염에 의한 변이주 특이성 단백질의 손실)

  • Ahn, Tae-In;Park, Eui-Yul
    • The Korean Journal of Zoology
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    • v.28 no.1
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    • pp.21-30
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    • 1985
  • By two-dimensional gel electrophoresis loss of a cell-specific protein was detected in tD strain of Amoeba proteus that had been infected by symbiotic bacteria extracted from xD strain. In 50 days of experimental infection by induced phagocytosis the host amoeba lost the ability to synthesize the tD cell-specific protein even after removal of the infective bacteria and xD cell-specific protein by growing the amoebae at $27^\\circC$. By this time the host amoebae were obligately dependent on the bacteria. From these and other results (Lorch and Jeon, Science 221:549), it is clear that the incompatibility of the infected nuclei with the cytoplasm of the uninfected amoeba and the obligate dependence of the host on bacteria are due to the irreversible inactivation or the loss of the cell-specific gene by bacterial infection in this amoeba.

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Characteristics of Zeolites (Zeolite의 특성)

  • Im, Goeng
    • The Journal of Natural Sciences
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    • v.6 no.1
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    • pp.103-108
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    • 1993
  • Zeolites were discovered as a natural mineral more than two hundred ago. In the beginning, the mineral was used as ion-exchange material and adsorbent. After the end of World War II , however, a variety of zeolites have become available in large amounts because of the establishment of low temperature synthesis and the discobery of natural zeolite deposits of sedimentary origin. Various uses of xeolite were developed utilizing the unique crystal strucrure and function of these minerals. In connection with this development remakable progress has also been made in basic stuides on the related geology and mineralogy, crystallization from sols and gels, structure, ion exchange, adsorption and cataiysis. As a result, zeolites, which had been known only as mineral specimens displayed in museums. established a firm position among the high-technology masterials with excellent functional capabilities.

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Improved Viability and Proteome Analysis of Lactobacillus fermentum KLB12 upon Heat Stress (Lactobacillus fermentum KLB12의 열 전처리에 따른 열 스트레스 내성 증진 및 프로테옴 변화)

  • 김주현;박미영;김승철;윤현식;소재성
    • KSBB Journal
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    • v.18 no.4
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    • pp.294-300
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    • 2003
  • In the previous study, we have isolated several vaginal lactobacilli from Korean woman and selected one of them (KLB12) for further study, which was indentified as Lactobacillus fermentum by sequence analysis of 16S rRNA gene. Formulated L. fermentum KLB12 can be used for ecological treatment of bacterial vaginosis. For pharmaceutical formulation, the spray-drying process is required where stress such as high temperature is routinely applied. In this study, we found that heat stress at 60$^{\circ}C$ for 20∼30min reduced the viable cell population of KLB12 by 10$\sub$6/~10$\sub$9/. However, adaptation of KLB12 cells at 52$^{\circ}C$ made them more thermotolerant upon exposure to 60$^{\circ}C$. The level of thermal protection in RSM (reconstituted skim milk) by adaptation in acid (pH 5), cold (4$^{\circ}C$), ethanol (3%), NaCI (0.3M) was also examined. Although not as efficient as the homologous stress, adaptations in both cold and NaCI gave considerable cross protection against heat stress. When chloramphenicol was added during heat adaptation, adaptation effect was abolished. This suggests that de novo protein synthesis is necessary during the adaptation process. Important changes in proteome during heat adaptation was examined with two-dimensional gel electrophoresis.