• Title/Summary/Keyword: cell-free system

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Optimization of Programmed Suppression in a Cell-Free Protein Synthesis System with Unnatural Amino Acid S-(2-Nitrobenzyl)cysteine

  • HYUN JOO;KANG, TAEK JIN;HUI KYOUNG SONG;JIN HO AHN;CHA YONG CHOI
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.344-347
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    • 2003
  • Unnatural amino acid S-(2-nitrobenzyl)cysteine was incorporated into human erythropoietin by using a programmed suppression of nonsense codon in a cell-free protein synthesis system. Several controlling factors affecting the operational efficiency of the suppression were investigated and optimized. The amount of suppressor tRNA and the concentration of $Mg^2+$ were crucial not only for the efficiency but also for the control of the exact suppression. In addition, some general optimization factor are reported in order to improve the efficiency in an unnatural amino acid mutagenesis.

Energy-efficient mmWave cell-free massive MIMO downlink transmission with low-resolution DACs and phase shifters

  • Seung-Eun Hong;Jee-Hyeon Na
    • ETRI Journal
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    • v.44 no.6
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    • pp.885-902
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    • 2022
  • The mmWave cell-free massive MIMO (CFmMIMO), combining the advantages of wide bandwidth in the mmWave frequency band and the high- and uniform-spectral efficiency of CFmMIMO, has recently emerged as one of the enabling technologies for 6G. In this paper, we propose a novel framework for energy-efficient mmWave CFmMIMO systems that uses low-resolution digital-analog converters (DACs) and phase shifters (PSs) to introduce lowcomplexity hybrid precoding. Additionally, we propose a heuristic pilot allocation scheme that makes the best effort to slash some interference from copilot users. The simulation results show that the proposed hybrid precoding and pilot allocation scheme outperforms the existing schemes. Furthermore, we reveal the relationship between the energy and spectral efficiencies for the proposed mmWave CFmMIMO system by modeling the whole network power consumption and observe that the introduction of low-resolution DACs and PSs is effective in increasing the energy efficiency by compromising the spectral efficiency and the network power consumption.

Calcium Ion Effect on the Sugar-H+ -Cotransport System in Chlorella vulgaris (Chlorella vulgaris의 당류 능동수송계에 미치는 칼슘 이온의 영향)

  • 조봉희
    • Journal of Plant Biology
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    • v.36 no.4
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    • pp.321-326
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    • 1993
  • Sugar uptake is accompanied with H+-substrate-symport generally. Both H+/sugar-and H+/K+ stoichiometries during the sugar-uptake have been reported to be exactly 1 : 1. This paper reports that the stoichiometries were enhanced dramatically by the addition of CaCl2 into the medium and by the high cell density of 200 $\mu$L pc/mL. The concentration of free Ca2+ ions in the cells increased significantly with cell density. It is suggested that the free Ca2+ ions are responsible for the change of stoichiometry of sugar transport system by regulation of H+ ion level of biomembrane.

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Use of Moving Aeration Membrane Bioreactor for the Efficient Production of Tissue Type Plasminogen Activator in Serum Free Medium

  • Hyun Koo Kim;Moo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.1 no.1
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    • pp.32-35
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    • 1996
  • Amoving aeration-membrane (MAM) bioreactor was employed for the production of 2$\mu$g/mL of tissue type Plasminogen Activator (tPA)in serum free medium from normal human fibroblast cells. This system could maintain high cell density for long periods of steady state conditions in perfusion cultivation. Under normal operating condition, shear stress was as low as 0.65 dynes/$\textrm{cm}^2$ at the agitation speed of 80 rpm. Even though cell density gradually decreased with increasing agitation speed, tPA production increased linearly with increasing shear stress within a moderate range. This culture system allowed production of 2$\mu$g tPA/mL while maintaining a high cell denisty of 1.0$\times$107 viable cell/mL.

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Enhanced Synthesis of Active rPA in the Continuous Exchange Cell-free Protein Synthesis [CECF] System utilizing Molecular Chaperones (분자 샤페론을 사용한 연속확산식 무세포단백질 발현 시스템에서의 재조합 Plasminogen Activator의 효율적 발현)

  • Park, Chang-Gil;Kim, Tae-Wan;Choi, Cha-Yong;Kim, Dong-Myung
    • KSBB Journal
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    • v.21 no.2
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    • pp.118-122
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    • 2006
  • In this report, we describe that the use of GroEL/GroES-enriched S30 extract remarkably enhances the solubility and enzymatic activity of cell-free synthesized rPA, which requires the correct formation of 9 disulfide bonds for its biological activity. We found that the stable maintenance of redox potential is necessary, but not sufficient for the optimal expression of active rPA. In a control reaction without using additional molecular chaperones, most of the rPA molecules were aggregated almost instantly after their expression and thus failed to exhibit the enzymatic activity. However, by the use of GroEL/GroES-enriched extract, combined with IAM-treatment, approximately $30{\mu}g/ml$ of active rPA was expressed in the cell-free synthesis reaction. This result not only demonstrates the efficient production of complex proteins, but also shows the control and flexibility offered by the cell-free protein synthesis system.

Design of Denitrification Reactor by Using Permeabilized and Immobilized Paracoccus denitrificans (Permeabilized Paracoccus denitrificans를 이용한 고정화 균주의 탈질화 반응기 설계)

  • Yun, Mi-Sun;Song, Ju-Yeong;Park, Keun-Ho
    • KSBB Journal
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    • v.20 no.2 s.91
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    • pp.100-105
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    • 2005
  • Removal of nitrogen compound from waste water is essential and often accomplished by biological process. Denitrification bacterium, Paracoccus denitrificans (KCTC 2350) is employed to estimate the denitrification ability and the characteristics. In the immobilized biological reactor system, the measurement of absolute amount of active strain in the reactor is comparatively difficult or impossible. In this. study, a reactor was designed with the unwoven texture wrapped peep holed plastic tube to calculate the absolute amount of active strain by comparing the activity of the permeabilized and or immobilized reactor and the free cell reactor The reactor system was continuous stirred tank reactor and the reaction rate of substrate consumption was assumed to satisfy the Michaelis-Menten equation. The effluent concentration of nitrate and nitrite was measured to estimate the apparent parameter of Michaelis-Menten equation. As a result, we found that the amount of immobilized active strain was figured out to be half of the total active strain in the reactor and the time required to be reached in the equilibrium state in the permeabilized and or immobilized reactor system was figured out to be shorter than that of the free cell reactor system.

Modeling of selfie stick using FreeCAD (FreeCAD를 이용한 셀카봉 모델링)

  • Sung, Gwangwon;Kim, Youngki
    • Proceeding of EDISON Challenge
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    • 2015.03a
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    • pp.428-441
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    • 2015
  • Corresponding to the high consumption rate of the selfie stick for cell phone camera, the design of the product is becoming diverse. However, relatively low cost selfie stick models are not strong enough to stand the weight of the cell phone. This results into a bending of the stick or the accidental fall of the cell phone by the failure of the holding unit. As a solution to the problem, design of selfie stick that is structurally strong enough to stand the weight of the cell phone with the minimum weight for the portability is proposed in this paper.

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Effects of Synthetic Pseudoceramides on Sphingosine Kinase Activity in F9-12 Cells

  • Jin, You-Xun;Shin, Kyong-Oh;Park, Myung-Yong;Lee, Shin-Hee;Park, Byeong-Deog;Oh, Sei-Kwan;Yoo, Hwan-Soo;Lee, Yong-Moon
    • Biomolecules & Therapeutics
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    • v.19 no.1
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    • pp.134-139
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    • 2011
  • Sphingosine kinase (SPHK) has a central role to control cell death and cell proliferation, which is suggested as a sphingolipid rheostat by regulating the levels between ceramide and sphingosine 1-phosphate (S1P). Therefore, physiological regulators of SPHK will be a good candidate to develop a new targeted drug. For this purpose, a series of synthetic pseudoceramides were tested by SPHK assay either cell-based or cell-free system. K10PC-5 strongly inhibited SPHK, while K6PC-5 activated SPHK in cell-free system. Specifically, K6PC-5 activated SPHK under the co-treatment with $50\;{\mu}M$ dimethylsphingosine (DMS), a SPHK inhibitor. Collectively, we developed a simple SPHK assay system to find SPHK regulatory pseudoceramide compounds, K10PC-5 and K6PC-5 which may be useful to cancer treatment or immune regulation like FTY720, a synthetic sphingolipid mimetic compound.

Tetramethylpyrazine Production by Immobilized Culture of Lactococcus lactis subsp. lactis biovar. diacetilactis FCl

  • Lee, Ji-Eun;Woo, Gun-Jo;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.6 no.2
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    • pp.137-141
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    • 1996
  • Acetoin and ammonia, the precursors of tetramethylpyrazine (TMP) having "meaty" or "roasted" flavors, were produced by the culture of Lactococcus lactis ssp. lactis biovar. diacetilactis FC1 in free and immobilized cell systems. Cells were immobilized using k-carrageenan and then were incubated at $34^{\circ}C$. The TMP productivity (0.34 g/l) and the conversion ratio (9.3%) of acetoin to TMP of the immobilized cell system were higher than those (0.24 g/l, 7.0%) of the free cell system. When the beads were activated for 12 h, the productivity of acetoin and TMP increased slightly.

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Differential Expression of Glucose Transporter Gene in Mouse Early Embryos (생쥐 초기배아의 Glucose Transporter유전자 발현 양상에 관한 연구)

  • Youm, Hye-Won;Byun, Hye-Kyung;Song, Gyun-Ji;Kim, Hae-Kwon;Lee, Ho-Joon
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.1
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    • pp.77-86
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    • 1998
  • The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose-transporter (GLUT) proteins. The aim of this study was to determine which class of glucose transporter molecules was responsible for uptake of glucose in the mouse early embryo and at which stage the corresponding genes were expressed. In addition, co-culture system with vero cell was used to investigate the effect of the system on GLUT expression. Two-cell stage embryos were collected from the superovulated ICR female and divided into 3 groups. As a control, embryos were cultured in 0.4% BSA-T6 medium which includes glucose. For the experimental groups, embryos were cultured in either co-culture system with vero cells or glucose-free T6 medium supplemented with 0.4% BSA and pyruvate as an energy substrate. 2-cell to blastocyst stage embryos in those groups were respectively collected into microtubes (50 embryos/tube). Total RNA was extracted and RT-PCR was performed. The products were analysed after staining ethidium bromide by 2% agarose gel electrophoresis. Blastocysts were collected from each group at l20hr after hCG injection. They were fixed in 2.5% glutaraldehyde, stained with hoechst, and mounted for observation. In control, GLUT1 was expressed from 4-cell to blastocyst. GLUT2 and GLUT3 were expressed in morula and blastocyst. GLUT4 was expressed in all stages. When embryos were cultured in glucose-free medium, no significant difference was shown in the expression of GLUT1, 2 and 3, compared to control. However GLUT4 was not expressed until morular stage. When embryos were co-cultured with vero cell, there was no significant difference in the expression of GLUT1, 2, 3 and 4 compared to control. To determine cell growth of embryos, the average cell number of blastocyst was counted. The cell number of co-culture ($93.8{\pm}3.1$, n=35) is significantly higher than that of control and glucose-free group ($76.6{\pm}3.8$, n=35 and $68.2{\pm}4.3$, n=30). This study shows that the GLUT genes are expressed differently according to embryo stage. GLUTs were detectable throughout mouse preimplantation development in control and co-culture groups. However, GLUT4 was not detected from 2- to 8-cell stage but detected from morula stage in glucose-free medium, suggested that GLUT genes are expressed autocrinally in the embryo regardless of the presence of glucose as an energy substrate. In addition, co-culture system can increase the cell count of blastocyst but not improve the expression of GLUT. In conclusion, expression of GLUT is dependent on embryo stage in preimplantation embryo development.

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