• Title/Summary/Keyword: shuttle mechanism

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Effects of amino acids on ethanol metabolism and oxidative stress in the ethanol-perfused rat liver

  • Park, Yeong-Chul;Oh, Se-In;Lee, Mee-Sook;Park, Sang-Chul
    • Environmental Mutagens and Carcinogens
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    • v.16 no.1
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    • pp.13-18
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    • 1996
  • One mechanism of free-radical production by ethanol is suggested to be through the intracellular conversion of XDH to XO by increased ratio of NADH to NAD. The major mechanism for physiological compensation of cytosolic NADH/NAD balance is the malate/aspartate shutfie. Therefore, it is important to develop the method to improve the efficiency of malate/aspartate shuttle in ethanol metabolism. In the present study, various amino acids and organic acid involved in the shuttle were tested for their functional efficiency in modulating shuttle in the ethanol-perfused rat liver. The rate of ethanol oxidation in the liver perfused with aspartate alone or aspartate in combination with pyruvate, respectively, was increased by about 10% compared to control liver, but not in the tissues perfused with glummate, cysteine or pyruvate alone. Though glummate, cysteine and pyravate did not affect the ethanol oxidation significanfiy, they showed some suppresive effect on the ethanol-induced radical generation monitored by protein carbonylation analysis. Among the tested components, aspartate is confirmed to be the most efficient as a metabolic regulator for both ethanol oxidation and ethanol-induced oxidative stress in our perfusion system. These effects of aspartate would result from NAD recycling by its supplementation through the coupled aspartate aminotransferase/malate dehydrogenase reactions and the malate-aspartate shuttle.

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Studies on the $HIS_5$ Gene of Yeast -$HIS_5-lacZ$ fusion and regulation in Saccharomyces cerevisiae- (효모 $HIS_5$ 유전자에 관한 연구 -Saccharomyces cerevisiae의 $HIS_5-lacZ$ 융합과 조절-)

  • Chung, Dong-Hyo;Oshima, Yasuji;Nishiwaki, Kyoni
    • Applied Biological Chemistry
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    • v.28 no.1
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    • pp.36-47
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    • 1985
  • $HIS_5$ gene of Saccaromyces cerevisiae was cloned into pBR 322 and also into pSH 610 shuttle vector. $HIS_5$ gene was expressed as promoter of lactose operon(lacZ). And $HIS_5-lacZ$ fusion was intergrated into chromosome III of yeast. $HIS_5$ gene in yeast growth was controlled general amino acid control mechanism.

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Molecular Characterization of Plasmid from Bifidobacterium longum

  • Park, Myeong-Soo;Moon, Hye-Won;Ji, Geun-Eog
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.457-462
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    • 2003
  • The complete nucleotide sequence of a plasmid, pMG1, isolated from Bifidobacterium longum MG1 has been determined. This plasmid, composed of 3,862 base pairs with 65.1% of G+C content. harbors two major open reading frames (ORF) encoding putative proteins of 29 kDa (ORF I) and 71 kDa (ORF II). ORF I showed relatively high amino acid sequence homology with replication proteins of other plasmids from Gr Im-positive and -negative bacteria. Upstream of ORF I, four sets of tandem repeat sequences resembling the iteron structure of related plasmids were found. S1 endonuclease treatment and Southern blot analysis revealed that pMG1 accumulates single-stranded DNA (ssDNA) intermediate, which indicate i the rolling circle replication (RCR) mechanism of this plasmid. Homology search indicated that ORF II encodes plasmid mobilization protein, and the presence of highly conserved oriT sequence in the upstream of this gene supported this assumption. RT-PCR showed that only ORF I is expressed in vivo. Based on these results, pMG 1 was exploited to construct a shuttle vector, pBES2. It was successfully transformed into Bifidobacterium and maintained stably.

Characterization of Plasmid pKJ36 from Bifidobacterium longum and Construction of an E. coli-Bifidobacterium Shuttle Vector

  • Park, Nyeong-Soo;Shin, Dong-Woo;Lee, Ke-Ho;Ji, Geun-Eog
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.312-320
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    • 2000
  • Abstract The full sequence of the plasmid pKJ36, which was derived from Bifidobacterium longum KJ, was determined and analyzed to construct shuttle vectors between E. coli and Bifidobacterium. The plasmid pKJ36 was composed of 3,625 base pairs with a 65.1% G+C content. The structural organization of pKJ36 was highly similar to that of pKJ50, and the three major ORFs on pKJ36 showed high amino acid sequence homologies with those of pKJ50. The putative proteins coded by these three ORFs were designated as RepB (32.0 kDa, pI=9.25), MembB (29.0 kDa, pI=12.25), and MobB (39.0 kDa, pI=IO.66), respectively. The amino acid sequence of RepB showed a 57% identity and 70% similarity with that of the RepA protein of pKJ50. Upstream of the repB gene, the so-called iteron sequence was directly repeated four-and-ahalf times and a conserved dnaA box was identified. An amino acid sequence comparison between the MobB and MobA of pKJ50 revealed a 48% identity and 61 % similarity. A conserved oriT sequence with an inverted repeat identical to that of pKJ50 was also found upstream of the mobB gene. A hydropathy analysis of MembB revealed four possible transmembrane regions. The expressions of the repB and membB genes were confirmed by RT-PCR. The in vitro translation reaction of pKJ36 showed protein bands with anticipated sizes with respect to each putative gene product. S 1 endonuclease treatment and Southern hybridization suggested that pKJ36 replicates by a rolling circle mechanism via a single-stranded DNA (ssDNA) intermediate. A shuttle vector between E. coli and Bifidobacterium sp. was constructed using the pKJ36, pBR322, and staphylococcal chloramphenicol acetyl transferase (CAT) gene. The successful transformation of the Bifidobacterium strains was shown by Southern hybridization and PCR. The transformation efficiency differed from strain to strain and, depending on the electroporation conditions, with a range between $1.2{\times}10^1-2.6{\times}10^2{\;}cfu/\mu\textrm{g}$ DNA.X> DNA.

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Proposal of Pipe Pressure Mode Analysis Method in Propulsion System for Predicting the Pogo of Space Launch Vehicle (우주 발사체의 포고현상 예측을 위한 공급/추진계의 파이프 압력모드 해석 기법 제안)

  • Lee, SangGu;Lee, SiHun;Shin, SangJoon
    • Proceedings of the Korean Society of Propulsion Engineers Conference
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    • 2017.05a
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    • pp.714-717
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    • 2017
  • Among the factors considered in the design stage of a space launch vehicle using liquid propellant, research has been focused out on the pogo phenomenon, longitudinal dynamic instability. The pogo phenomenon refers to the instability that the longitudinal vibration of the launch vehicle structure causes a change in the pressure and flow rate of the fluids in propulsion system, and this change re-excites the fuselage structure. This mechanism constitutes a closed system to gradually increase the vibration of the launch vehicle. This paper specifically focuses on the dynamic analysis of pressure and flow changes in the propulsion system. Based on the example study of the space shuttle, the acoustic modal analysis of the propulsion system is performed to predict the modes of the supply line causing instability of the fuselage.

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Reentry Guidance for Korean Space Plane Based on Reference Drag Following (한국형 우주비행기의 기준 항력 추종 기반 재진입 유도 기법)

  • Yoon, Da-In;Kim, Young-Won;Lee, Chang-Hun;Choi, Han-Lim;Ryu, Hyeok
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.49 no.8
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    • pp.637-648
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    • 2021
  • This paper aims to propose new reentry guidance for Korean Space plane (KSP). Similar to the Space Shuttle guidance concept, a reference drag profile is first determined to satisfy several flight path constraints and boundary conditions, and the proposed guidance commands are realized in a way to track the predetermined reference drag profile. To this end, the drag dynamics is examined. The investigation uncovers that the dynamics characteristics of the drag and the flight path angle are considerably different. Based on this fact, the proposed guidance commands are determined using the time-scale separation technique and the feedback linearization methodology. The key feature of the proposed guidance lies in its simple structure and a clear working mechanism. Therefore, the proposed method is simple to implement compared to existing methods. Numerical simulations are performed to investigate the performance of the proposed method.

Transgenic Mutagenesis Assay to Elucidaate the Mechanism of Mutation at Gene Level (유전자수준에서 돌연변이 유발기전을 밝히는 Transgenic Mutagenesis Assay)

  • Ryu, Jae-Chun;Youn, Ji-Youn;Cho, Kyung-Hae;Chang, Il-Moo
    • Environmental Mutagens and Carcinogens
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    • v.18 no.1
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    • pp.15-21
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    • 1998
  • Transgenic animal and cell line models which are recently developed and used in toxicology fields combined with molecular biological technique, are powerful tools to study the mechanism of mutation in vivo and in vitro, respectively. Transgenic models, which have exogenous DNA incorporated into their genome, carry recoverable shuttle vector containing reporter genes to assess endogenous effects or alteration in specific genes related to disease processes. The lac I and lac Z gnee most widely used as a mutational target in transgenic systems. The assay is performed by treatment with putative mutagenic agents, isolation of genomic DNA from cells or tissues, exposure the isolated DNA to in vitro packaging extract, plating and sequencing. The results from these processes provide not only mutant frequency as quantitative evaluation but also mutational spectrum as qualitative evaluation of various agents. Therefore we introduce and review the principle, detailed procedure and application of transgenic mutagenesis assay system in toxicology fields especially in mutagenesis and carcinogenesis.

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Precision Position Controller of Linear Motor-Based Container Transfer System (선형전동기 기반 컨테이너 이송 시스템의 정밀 위치제어)

  • Lee, Young-Jin;Lee, Jin-Woo;Cho, Hyun-Cheol;Lee, Kwon-Soon
    • The Transactions of the Korean Institute of Electrical Engineers P
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    • v.57 no.3
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    • pp.215-224
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    • 2008
  • In this paper, we introduced a linear motor-based transfer system with an active pid controller which can be replaced with an automated guided vehicle (AGV) for the port automation. This system, which is named LMCTS(liner motor-based container transfer system), is based on PMLSM (permanent magnetic linear synchronous motor) which basically consists of stator modules on the rail and shuttle car. Therefore more progressive and adaptive control mechanisms should be required to control a system with large variation of container weight, the difference of each characteristic of stator modules, a stator module's trouble etc. We introduced an active control mechanism with an online tuning scheme using modified evolutionary strategy. Some computer simulations are implemented to assess the robustness of the proposed system.

Isolation and Characterization of a Cryptic Plasmid, pMBLR00, from Leuconostoc mesenteroides subsp. mesenteroides KCTC 3733

  • Chae, Han Seung;Lee, Jeong Min;Lee, Ju-Hoon;Lee, Pyung Cheon
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.837-842
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    • 2013
  • A cryptic plasmid, pMBLR00, from Leuconostoc mesenteroides subsp. mesenteroides KCTC 3733 was isolated, characterized, and used for the construction of a cloning vector to engineer Leuconostoc species. pMBLR00 is a rolling circle replication plasmid, containing 3,370 base pairs. Sequence analysis revealed that pMBLR00 has 3 open reading frames: Cop (copy number control protein), Rep (replication protein), and Mob (mobilization protein). pMBLR00 replicates by rolling circle replication, which was confirmed by the presence of a conserved double-stranded origin and single-stranded DNA intermediates. An Escherichia coli-Leuconostoc shuttle vector, pMBLR02, was constructed and was able to replicate in Leuconostoc citreum 95. pMBLR02 could be a useful genetic tool for metabolic engineering and the genetic study of Leuconostoc species.

Study on preparation of precipitated calcium carbonate using recycling water of ready-mixed Concrete (레미콘 회수수를 이용한 침강성 탄산칼슘 제조에 관한 연구)

  • Shin, Jae Ran;Kim, Jae Gang;Kim, Hae Gi;Kang, Ho Jong
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.2
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    • pp.232-238
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    • 2016
  • In this study, a liquid carbonation method was applied for producing precipitate calcium carbonate by liquid-liquid reaction. Also a shuttle mechanism of wet chemical absorption using MEA was utilized. The high concentration $CO_2$(A) and exhaust gas(B) was used for collecting carbon dioxide in the 30% MEA aqueous solution, and $CO_2$ was fixed with rate of 0.35 mg of $CO_2$ per mg of sludge through the liquid carbonation process. It was found from SEM data that calcium carbonate was mainly made up with spherical vaerite with the mixing of a small quantity of calcite.