• Title/Summary/Keyword: Repressing

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Variations in Protein Glycosylation in Hansenula polymorpha Depending on Cell Culture Stage

  • Kim, So-Young;Sohn, Jung-Hoon;Pyun, Yu-Ryang;Choi, Eui-Sung
    • Journal of Microbiology and Biotechnology
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    • v.17 no.12
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    • pp.1949-1954
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    • 2007
  • A simple way to prevent protein hyperglycosylation in Hansenula polymorpha was found. When glucose oxidase from Aspergillus niger and carboxymethyl cellulase from Bacillus subtilis were expressed under the control of an inducible methanol oxidase (MOX) promoter using methanol as a carbon source, hyperglycosylated forms occurred. In contrast, MOX-repressing carbon sources (e.g., glucose, sorbitol, and glycerol) greatly reduced the extent of hyperglycosylation. Carbon source starvation of the cells also reduced the level of glycosylation, which was reversed to hyperglycosylation by the resumption of cell growth. It was concluded that the proteins expressed under actively growing conditions are produced as hyperglycosylated forms, whereas those under slow or nongrowing conditions are as short-glycosylated forms. The prevention of hyperglycosylation in the Hansenula polymorpha expression system constitutes an additional advantage over the traditional Saccharomyces cerevisiae system in recombinant production of glycosylated proteins.

Paclitaxel Stimulates Cyclooxygenase-2 Expression via MAP Kinase Pathway in Rabbit Articular Chondrocytes

  • Im, Jeong-Hee;Kim, Song-Ja
    • Biomedical Science Letters
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    • v.15 no.2
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    • pp.141-146
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    • 2009
  • Paclitaxel, an antimicrotubule agent, binds to beta-tubulin in the microtubule and stabilizes the polymer, thereby repressing dynamic instability. Here, we have demonstrated that microtubule cytoskeletal architecture involved in regulation of the COX-2 expression in chondrocyte treated with paclitaxel. Paclitaxel enhanced COX-2 expression and prostaglandin E2 production, as indicated by the Western blot analysis, reverse transcriptase PCR(RT-PCR) and immunofluorescence staining, and $PGE_2$ assay, respectively. In our previous data have shown that paclitaxel treatment stimulated activation of ERK-1/2 and p38 kinase(Im et al., 2009). SB203580, an inhibitor of p38 kinase, blocked the induction of COX-2 expression by paclitaxel. Also PD98059, an inhibitor of ERK-1/2 kinase was blocked the induced COX-2 expression. These results indicate that activation of ERK-1/2 and p38 kinase is required for COX-2 expression induced by paclitaxel in rabbit articular chondrocytes.

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Inhibition of methane formation and improvement of $H_2$ production by adding nitrate to anaerobic reactor

  • Kim, Jeong-Ok;Kim, Yong-Hwan;Ryu, Jeong-Yong;Song, Bong-Keun;Kim, In-Ho
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.129-134
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    • 2003
  • Anaerobic hydrogen production was executed using mixed anaerobic sludge. pH control (pH $5.5^{\sim}6.2)$ and heat treatment of anaerobic sluge at $92^{\circ}C$ was not effective for repressing the methane formation. The addition of $nitrate(500^{\sim}2,000\;mg/L\;KNO_3)$ made it possible to repress the methane formation, which resulted in the improvement of hydrogen production. By using SEM(Scanning Electron Microscope), less methane forming microbial with spagetti shape was observed when nitrate was supplied to anaerobic reactor.

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Modulation of Phosphoenolpyruvate Metabolism of Anaerobiospirillum succiniciproducens ATCC 29305

  • Yoo, Jin Young;J. Gregory Zeikus
    • Journal of Microbiology and Biotechnology
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    • v.6 no.1
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    • pp.43-49
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    • 1996
  • Modulation of the catabolic PEP-pathway of Anaerobiospirillum succiniciproducens was tried using some enzymatic inhibitors such as gases and chemicals in order to enhance succinic acid production. 10$\%$ CO increased the succinic acid/acetic acid (S/A) ratio but inhibited growth as well as production of succinic and acetic acid. Hydrogen gas also increased the S/A ratio and inhibited the synthesis of pyruvate: ferredoxin oxidoreductase when used in mixture with $CO_2$, Catabolic repression by acetic, lactic and formic acid was not recognized and other modulators such as glyoxylate, pyruvate derivatives, arsenic salt, phosphate and sulfate were shown not to be effective. Magesium carbonate was shown effective for repressing acetate production. Palmitic acid, myristic acid and phenylalanine did not affect acetate production but carprylic acid completely inhibited growth.

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Anti-metastatic mechanism of mountain cultivated wild ginseng in human cancer cell line

  • Jang, S.B.;Lim, C.S.;Jang, J.H.;Kwon, K.R.
    • Journal of Pharmacopuncture
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    • v.13 no.1
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    • pp.37-43
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    • 2010
  • Objective : Ginseng is one of most widely used herbal medicine. Ginseng showed anti-metastasis activities. However, its molecular mechanisms of action are unknown. So we want to report the wild ginseng repress which plays key roles in neoplastic epithelial-mesenchymal transition process. Methods : Treatment of the human colorectal carcinoma LOVO cells and human gastric carcinoma SNU601 cells with the increased concentrations of cultivated wild ginseng extracts resulted in a gradual decrease in the AXIN2 gene expression. Results : Metastasis-suppressor genes, maspin and nm23 was not affected by the treatment of ginseng extracts in LOVO cells. Moreover, the mountain cultivated wild ginseng or mountain wild ginseng are similar in their inhibitory effects on the expression of AXIN2 gene, but are substantially stronger than cultivated ginseng. Conclusion : We described the novel mechanism of wild ginseng-induced anti-metastasis activity by repressing the expression of AXIN2 gene that plays key roles in epithelial-mesenchymal transition process.

Sequence Analysis and Functional Expression of the Structural and ]Regulatory Genes for Pyruvate Dehydrogenase

  • Hwan Youn;Jangyul Kwak
    • Journal of Microbiology
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    • v.40 no.1
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    • pp.43-50
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    • 2002
  • A cluster of genes encoding the pyruvate dehydrogenase complex (PDC) of Streptomyces seoulensis, a Gram-positive bacterium, was cloned and sequenced. The genes of S. seoulensis consist of four open reading frames. The first gene, lpd, which encodes a lipoamide dehydrogenase, is followed by pdhB encoding a dihydrolipoamide acetyltransferase (E2p), pdhR, a regulatory gene, and pdhA encoding a pyruvate dehydrogenase component (Elp). Elp had an unusual homodimeric subunit, which has been known only in Gram-negative bacteria S. seoulensis E2p contains two lipoyl domains like those of humans and Streptomyces faecalis. The pdhR gene appears to be clustered with the structural genes of S. seoulensis PDC. The PdhR-overexpressed S. seoulensis howed growth retardation and the decrease of Elp, indicating that PdhR regulates the function of PDC by repressing the expression of Elp. A strain of Streptomyces licidans overexpressing S. seoulensis PdhR showed a significant decreasein the level of actinorhodin, implying a regulatory role for Streptomyces PDC in antibiotic biosynthesis.

A study on the design of the torsional vibration viscous damper for the crankshaft and developing of its performance simulation computer program (크랭크축 비틀림진동점성댐퍼의 설계와 댐퍼 성능시뮬레이션프로그램개발)

  • 이충기;전효중
    • Journal of Advanced Marine Engineering and Technology
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    • v.13 no.1
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    • pp.77-96
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    • 1989
  • In diesel engines, it is inevitable that the torsional vibration is produced by the fluctuation of engine torque. Therefore, if the occurence of torsional vibration is confirmed in the design stage or the torsional vibration is observed on the bed of test run, it is necessary to establish some preventive measures to avoid dangerous conditions. Major preventive measures are as follows : 1. Changing the natural frequency of shaft system. 2. Repressing the vibration amplitude by the damping energy. 3. Counterbalancing the exciting torque by the resistant torque. 4. Counterbalacing the harmonic component of exciting energy. In above methos, the damper is the last measure to be used for controlling the torsional vibration. In this thesis, the design of viscous damper that absorbs the exciting energy is investigated and a number of problems associated with the design of viscous damper are treated and a computer pregram for the process of damper design is developed. A viscous damper for a high speed diesel engine is designed and its effect is simulated by the author's computer program.

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A study on the development of emergency management program for railway fire safety improvement (철도화재 안전개선대책수립을 위한 위기관리프로그램 개발 연구)

  • Wang, Jong-Bae;Kwak, Sang-Log;Hong, Seon-Ho
    • Proceedings of the KSR Conference
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    • 2003.10b
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    • pp.235-240
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    • 2003
  • In this paper, the fire safety management plan and the emergency action program for the railway safety improvement were reviewed. In the fire safety management plan, a series of preventive methods answering to such as a smoke exhaust installation, a fire warning device and a guide inducement will be come into preparation. Also, the damage reducing methods such as complement of fire safety equipments, establishment of refuge & escape system, consolidation of real-time control on fire response and operation procedure will be required for early repressing fire and minimizing fatality.

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Mesodermal repression of single-minded in Drosophila embryo is mediated by a cluster of Snail-binding sites proximal to the early promoter

  • Park, Kye-Won;Hong, Joung-Woo
    • BMB Reports
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    • v.45 no.10
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    • pp.577-582
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    • 2012
  • single-minded (sim) is a master regulatory gene that directs differentiation in the central nervous system during Drosophila embryogenesis. Recent identification of the mesectoderm enhancer (MSE) of sim has led to the hypothesis that two Snail (Sna)-binding sites in the MSE may repress sim expression in the presumptive mesoderm. We provide evidence here that three Sna-binding sites proximal to the sim promoter, but not those of the MSE, are responsible for the mesodermal repression of sim in vivo. Using transgenic embryos injected with lacZ transgenes, we showed that sim repression in the mesoderm requires the three promoter-proximal Sna-binding sites. These results suggest that Sna represses the mesectodermal expression of sim by directly repressing the nearby promoter, and not by quenching adjacent transcriptional activators in the MSE. These data also showed how the MSE, lacking the three proximal Sna-binding sites, reproduced the endogenous pattern of sim expression in transgenic embryos.

Multiple shRNA expressing vector enhances efficiency of gene silencing

  • Song, Jun;Giang, An;Lu, Yingchun;Pang, Shen;Chiu, Robert
    • BMB Reports
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    • v.41 no.5
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    • pp.358-362
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    • 2008
  • RNA interference (RNAi) is the process of sequence-specific gene silencing. However, RNAi efficiency still needs to be improved for effective inhibition of target genes. We have developed an effective strategy to express multiple shRNAs (small hairpin RNA) simultaneously using multiple RNA Polymerase III (Pol III) promoters in a single vector. Our data demonstrate that multiple shRNAs expressed from Pol III promoters have a synergistic effect in repressing the target gene. Silencing of endogenous cyclophilin A (CypA) or key HIV viral genes by multiple shRNAs results in significant inhibition of the target gene.