• 제목/요약/키워드: two-dimensional gel

검색결과 277건 처리시간 0.022초

Metagenomic and Proteomic Analyses of a Mangrove Microbial Community Following Green Macroalgae Enteromorpha prolifera Degradation

  • Wu, Yijing;Zhao, Chao;Xiao, Zheng;Lin, Hetong;Ruan, Lingwei;Liu, Bin
    • Journal of Microbiology and Biotechnology
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    • 제26권12호
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    • pp.2127-2137
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    • 2016
  • A mangrove microbial community was analyzed at the gene and protein levels using metagenomic and proteomic methods with the green macroalgae Enteromorpha prolifera as the substrate. Total DNA was sequenced on the Illumina HiSeq 2000 PE-100 platform. Two-dimensional gel electrophoresis in combination with liquid chromatography tandem mass spectrometry was used for proteomic analysis. The metagenomic data revealed that the orders Pseudomonadales, Rhizobiales, and Sphingomonadales were the most prevalent in the mangrove microbial community. By monitoring changes at the functional level, proteomic analyses detected ATP synthase and transporter proteins, which were expressed mainly by members of the phyla Proteobacteria and Bacteroidetes. Members of the phylum Proteobacteria expressed a high number of sugar transporters and demonstrated specialized and efficient digestion of various glycans. A few glycoside hydrolases were detected in members of the phylum Firmicutes, which appeared to be the main cellulose-degrading bacteria. This is the first report of multiple "omics" analysis of E. prolifera degradation. These results support the fact that key enzymes of glycoside hydrolase family were expressed in large quantities, indicating the high metabolic activity of the community.

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

  • 김정동;황병국
    • 한국식물병리학회지
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    • 제11권1호
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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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Induction of Drought Stress Resistance by Multi-Functional PGPR Bacillus licheniformis K11 in Pepper

  • Lim, Jong-Hui;Kim, Sang-Dal
    • The Plant Pathology Journal
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    • 제29권2호
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    • pp.201-208
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    • 2013
  • Drought stress is one of the major yield affecting factor for pepper plant. The effects of PGPRs were analyzed in relation with drought resistance. The PGPRs inoculated pepper plants tolerate the drought stress and survived as compared to non-inoculated pepper plants that died after 15 days of drought stress. Variations in protein and RNA accumulation patterns of inoculated and non-inoculated pepper plants subjected to drought conditions for 10 days were confirmed by two dimensional polyacrylamide gel electrophoresis (2D-PAGE) and differential display PCR (DD-PCR), respectively. A total of six differentially expressed stress proteins were identified in the treated pepper plants by 2D-PAGE. Among the stress proteins, specific genes of Cadhn, VA, sHSP and CaPR-10 showed more than a 1.5-fold expressed in amount in B. licheniformis K11-treated drought pepper compared to untreated drought pepper. The changes in proteins and gene expression patterns were attributed to the B. licheniformis K11. Accordingly, auxin and ACC deaminase producing PGPR B. licheniformis K11 could reduce drought stress in drought affected regions without the need for overusing agrochemicals and chemical fertilizer. These results will contribute to the development of a microbial agent for organic farming by PGPR.

대장암 세포에서 5-FU(Fluorouracil)의 세포독성과 관련된 단백체 분석 (Proteomic Approach to the Cytotoxicity of 5-FU(Fluorouracil) in Colon Cancer Cells)

  • 이서영;송진수;노시훈;김근태;홍순선;김희준;권성원;박정일
    • 약학회지
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    • 제53권3호
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    • pp.145-150
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    • 2009
  • We evaluated cytotoxic effect based on the MTT assay and identified altered proteins in 5-FU(fluorouracil) treated HT29 cells using two-dimensional gel electrophoresis and MALDI-TOF/TOF-MS. As proteins inducing apoptosis, siah binding protein 1 and p47 protein isoform a were up-regulated and tumor protein translationally-controlled 1 was down-regulated by 5-FU treatment. And mannose 6 phosphate receptor binding protein 1 controls DNA mismatch repair system was increased. We suggest 5-FU promotes a cytotoxicity under the action of these proteins in colon cancer cells.

Ginsenoside-Rp1-induced apolipoprotein A-1 expression in the LoVo human colon cancer cell line

  • Kim, Mi-Yeon;Yoo, Byong Chul;Cho, Jae Youl
    • Journal of Ginseng Research
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    • 제38권4호
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    • pp.251-255
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    • 2014
  • Background: Ginsenoside Rp1 (G-Rp1) is a novel ginsenoside derived from ginsenoside Rk1. This compound was reported to have anticancer, anti-platelet, and anti-inflammatory activities. In this study, we examined the molecular target of the antiproliferative and proapoptotic activities of G-Rp1. Methods: To examine the effects of G-Rp1, cell proliferation assays, propidium iodine staining, proteomic analysis by two-dimensional gel electrophoresis, immunoblotting analysis, and a knockdown strategy were used. Results: G-Rp1 dose-dependently suppressed the proliferation of colorectal cancer LoVo cells and increased their apoptosis. G-Rp1 markedly upregulated the protein level of apolipoprotein (Apo)-A1 in LoVo, SNU-407, DLD-1, SNU-638, AGS, KPL-4, and SK-BR-3 cells. The knockdown of Apo-A1 by its small-interfering RNA increased the levels of cleaved poly(ADP-ribose) polymerase and p53 and diminished the proliferation of LoVo cells. Conclusion: These results suggest that G-Rp1 may act as an anticancer agent by strongly inhibiting cell proliferation and enhancing apoptosis through upregulation of Apo-A1.

Gene Expression study of human chromosomal aneuploid

  • 이수만
    • 한국생물정보학회:학술대회논문집
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    • 한국생물정보시스템생물학회 2006년도 Principles and Practice of Microarray for Biomedical Researchers
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    • pp.98-107
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    • 2006
  • Chromosomal copy number changes (aneuploidies) are common in human populations. The extra chromosome can affect gene expression by whole-genome level. By gene expression microarray analysis, we want to find aberrant gene expression due to aneuploidies in Klinefelter (+X) and Down syndrome (+21). We have analyzed the inactivation status of X-linked genes in Klinefelter Syndrome (KS) by using X-linked cDNA microarray and cSNP analysis. We analyzed the expression of 190 X-linked genes by cDNA microarray from the lymphocytes of five KS patients and five females (XX) with normal males (XY) controls. cDNA microarray experiments and cSNP analysis showed the differentially expressed genes were similar between KS and XX cases. To analyze the differential gene expressions in Down Syndrome (DS), Amniotic Fluid (AF)cells were collected from 12 pregnancies at $16{\sim}18$ weeks of gestation in DS (n=6) and normal (n=6) subjects. We also analysis AF cells for a DNA microarray system and compared the chip data with two dimensional protein gel analysis of amniotic fluid. Our data may provide the basis for a more systematic identification of biological markers of fetal DS, thus leading to an improved understanding of pathogenesis for fetal DS.

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핀-튜브형 흡착탑 해석시 입자간 물질전달 모델의 타당성 검증 (Validity of Inter-Particle Models for the Mass-Transfer Kinetics of a Fin-Tube-Type Adsorption Bed)

  • 안상혁;홍상우;권오경;정재동
    • 설비공학논문집
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    • 제25권12호
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    • pp.660-667
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    • 2013
  • This study presents a numerical investigation of the heat and mass transfer kinetics of a fin-tube-type adsorption bed using a two-dimensional numerical model with silica-gel and water as the adsorbent and refrigerant pair. The performance is strongly affected by the heat and mass transfer in the adsorption bed, but the details of the mass transfer kinetics remain unclear. The validity of inter-particle models used to simulate mass-transfer kinetics were examined, such as a constant pressure model and non-constant pressure model, and the valid ranges of the diffusion ratio for each model are proposed. The COP and SCP have been numerically calculated as the performance indexes according to the diffusion ratio. The constant pressure model, which is commonly used in previous research, was found to be valid only in a limited range of diffusion ratio.

The Physiological Role of CPR1 in Saccharomyces cerevisiae KNU5377 against Menadione Stress by Proteomics

  • Kim, Il-Sup;Yun, Hae-Sun;Kwak, Sun-Hye;Jin, Ing-Nyol
    • Journal of Microbiology
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    • 제45권4호
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    • pp.326-332
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    • 2007
  • In order to understand the functional role of CPRl in Saccharomyces cerevisiae KNU5377 with regard to its multi-tolerance characteristics against high temperatures, inorganic acids, and oxidative stress conditions, whole cellular proteins were analyzed via liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). This procedure was followed by two-dimensional (2-D) gel electrophoresis. Under menadione stress conditions, the 23 upregulated proteins were clearly identified only in the wild- type strain of KNU5377. Among the proteins, Sodl1p Tsa1p, Ahp1, Cpr1p, Cpr3, Ssb2p, and Hsp12p were identified as components of antioxidant systems or protein-folding related systems. The CPR1 protein could not be completely detected in the $cpr1{\Delta}$ mutant of KNU5377 and the other upregulated proteins in the wild-type strain evidenced a clear correlation with the results of immunoblot analysis. Moreover, a reduction in growth patterns (about 50%) could be observed in the $cpr1{\Delta}$ mutant, as compared with that of the wild-type strain under mild MD stress conditions. These results indicate that the upregulation of CPR1 may contribute to tolerance against MD as an inducer of oxidative stress.

The Impacts of Photoperiods on Hypothalamic Proteins in the Reproductive Activities of Golden Hamsters

  • Choi, Don-Chan;Han, Eun-Hee
    • 한국발생생물학회지:발생과생식
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    • 제14권3호
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    • pp.185-197
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    • 2010
  • The reproduction of animals is a means to maintain their species. The golden hamsters are seasonal breeders, and their reproduction is regulated by the photoperiod. The shifts of the sexual functions imply the changes of the protein expression, and those are reflected by the proteome. Thus the present study was to examine via two dimensional polyacrylamide gel electrophoresis (2d PAGE), the physiological changes and the alterations of protein expressions in hypothalamus upon the reproductive situation related to the pineal gland. Among the pineal intact animals, the reproductive functions were sustained in animals housed in long photoperiod (LP) but regressed in animals housed in short photoperiod. And those pinealectomized animals showed high sexual activities regardless of photoperiod. Ultimately they were branched into dichotomy, sexually active and inactive animals. Apart from the changes of physiological parameters upon the reproductive conditions, there were obvious differences in proteins extracted from the hypothalamus. The hypothalamus of LP animals presented high levels of enzymes which are involved in the production of energy, glycolysis and Krebs cycle. The increased energy might be related to the GnRH synthesis in hypothalamus and indirectly to the constant cell divisions in spermatogenesis. Taken together, the impacts of the photoperiodic information controlling reproduction could be observed through 2d PAGE. Therefore, the present results suggest the potential of biomarkers collectively to diagnose the fertility and the infertility by way of proteomics in organs with regard to the reproductive system, further could be applied to diagnose various diseases.

Proteomic Dissection of Abiotic Stress Response in Crop Plants

  • Alam, Iftekhar;Sharmin, Shamima Akhtar;Lee, Byung-Hyun
    • 한국환경농학회:학술대회논문집
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    • 한국환경농학회 2011년도 30주년 정기총회 및 국제심포지엄
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    • pp.196-204
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    • 2011
  • Abiotic stress is the primary cause of crop loss worldwide, reducing average yields for most major crop plants by more than 50%. In addition, future agricultural production and management will encounter multifaceted challenges from global climate change. Therefore, it is necessary to study the molecular response of crop plants to the stresses in order to develop appropriate strategies to sustain food production under adverse environmental conditions. We carried out a large scale proteomic analysis of soybean plants in response to various abiotic stresses, including drought, salinity, waterlogging and their interactions. Proteins were analyzed by two dimensional polyacrylamide gel electrophoresis followed by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry. The identified proteins are involved in a wide range of cellular functions. In addition to the well known stress-associated proteins, we identified several novel proteins, which were not reported before. In many cases our proteomic data bridges the gap between mRNA and metabolite data. Our studie provides new insights into identification of abiotic stress responsive proteins in soybean, and demonstrates the advantages of proteomic analysis in dissecting metabolic and regulatory networks.

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