• Title/Summary/Keyword: gel mobility

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Nucleotide Sequence and Inducibility Analysis of Chloramphenicol Acetyltransferase Gene from Staphylococcus aureus R-plasmid pSBK203 (Staphylococcus aureus에서 분리된 R-plasmid pSBK203상의 chloramphenicol acetyltransferase 인자의 염기서열 및 유발성 분석)

  • 권동현;변우현
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.194-200
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    • 1989
  • The nucleotide sequence of inducible chloramphenicol acetyl-transferase(CAT) gene isolated from a small plasmid pSBK203 of Staphylococcus aureus was determined. The base sequence shows that structural gene of pSBK203-CAT encodes a protein of 213 amino acids and has a leader region which encodes a short polypeptide of 9 amino-acids in its upstream. vertical bar /sup 35/S vertical bar-Methionine labelled CAT gene product in minicell showed almost same mobility with pC194-CAT of which molecular weight is 24Kdal on polyacrylamide gel electrophoresis. Predicted amino acid sequence of pSBK203-CAT has revealed a high degree of homology with the CATs of pC194 and pC221 than those of cat-86, Tn9 and proteus mirabilis PM13.

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Inhibitory Effect of Paeoniflorin on Fos-Jun-DNA Complex Formation and Stimulation of Apoptosis in HL-60 Cells

  • Kwon, Hae-Young;Kim, Kyoung-Su;Park, Se-Yeon;Lee, Dug-Keun;Yang, Chul-Hak
    • BMB Reports
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    • v.34 no.1
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    • pp.28-32
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    • 2001
  • The Fos-Jun heterodimers are part of the regulatory network of gene expression and nuclear proteins encoded by proto-oncogenes. The activation of Fos-Jun is important in the transmission of the tumor-promoting signal from the extracellular environment to the nuclear transcription mechanism. To search for the inhibitors of the Fos-Jun DNA complex formation, several natural products were screened and water-soluble paeoniflorin reduced the binding activity of the Fos-Jun heterodimer. This active compound was purified by silica gel column chromatography and HPLC. The electrophoresis mobility shift assay and reverse-phase HPLC test showed that paeoniflorin reduced the AP-l function. The cytotoxic effect of paeoniflorin was observed in HL-60. These results indicate that paeoniflorin blocks the Fos-Jun heterodimer-binding site of the AP-l DNA and it also has cytotoxic effects on human leukemia cell lines.

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Protective Effects of Hwansodan(Huanshao-dan) Water Extract in Serum Deprivation-induced Apoptosis of PC12 Cells (환소단이 영양혈청 결핍성 PC12 신경세포의 apoptosis에 미치는 영향)

  • 임준식;김명선;소홍섭;이지현;한상혁;허윤;박래길;문병순
    • The Journal of Korean Medicine
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    • v.21 no.4
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    • pp.64-72
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    • 2000
  • Objectives : This study was designed to investigate the neuroprotective effect of Hwansodan(Huanshao-dan) on the apoptosis induced by withdrawal of neurotrophic support. Methods : PCl2 pheochromocytoma cells have been used extensively as a model for studying the cellular and molecular effects of neuronal cells. The viability of cells was measured by MTT assay. We used DNA fragmentation and caspase 3-like protease activation assay. Results : The water extract of Hwansodan(Huanshao-dan) significantly showed protective effects on serum and glucose deprivation-induced apoptotic death. Hwansodan(Huanshao-dan) also prevents DNA fragmentation and caspase 3-like protease activation, representing typical neuronal apoptotic phenomena in PCl2 pheochromocytoma cells and induces tyrosine phosphorylation of proteins around 44 kDa, which was identified as ERK1 with electrophoretic gel mobility shift by Western blot. In addition, MAPK kinase(MEK) inhibitor PD98059 and Ras inactivator, ${\alpha}-hydroxyfarnesylphosphonic$ acid attenuated the neuroprotective effects of Hwansodan(Huanshao-dan) in serum-deprived PCl2 cells. Conclusions : These results indicate that Ras/MEK/ERK signaling pathway plays a key role in neuroprotective effects of Hwansodan(Huanshao-dan) in serum-deprived PCl2 cells. Taken together, we suggest the possibility that Hwansodan(Huanshao-dan) might provide a neurotrophic-like activity in PCl2 cells.

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The Roles of Tryptophan and Histidine Residues in the Catalytic Activities $\beta$-Cyclodextrin Glucanotransferase from Bacillus firmus var. alkalophilus

  • Shin, Hyun-Dong;Kim, Chan;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.1
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    • pp.62-69
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    • 1999
  • In order to investigate the critical amino acid residues involved in the catalytic activities of $\beta$-cyclodextrin glucanotransferase ($\beta$-CGTase) excreted by Bacillus firmus var. alkalophilus, the amino acid residues in $\beta$-CGTase were modified by various site-specific amino acid modifying reagents. The cyclizing and amylolytic activities of $\beta$-CGTase were all seriously reduced after treatment with Woodward's reagent K (WRK) modifying aspartic/glutamic acid, N-bromosuccinimde (NBS) modifying tryptophan, and diethylpyrocarbonate (DEPC) modifying histidine residues. The roles of tryptophan and histidine residues in $\beta$-CGTase were further investigated by measuring the protection effect of various substrates during chemical modification, comparing protein mobility in native and affinity polyacrylamide gel electrophoresis containing soluble starch, and comparing the $K_m$ and $V_{max}$ values of native and modified enzymes. Tryptophan residues were identified as affecting substrate-binding ability rather than influencing catalytic activities. On the other hand, histidine residues influenced catalytic ability rather than substrate-binding ability, plus histidine modification had an effect on shifting the optimum pH and pH stability.

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RNA Binding Specificities of Double-Stranded RNA Binding Protein (RBF) as an Inhibitor of PRK Kinase (PKR인산화효소 억제인자인 이중선RNA결합단백질 (RBF)의 RNA결합특이성)

  • 박희성;최장원
    • Journal of Life Science
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    • v.6 no.4
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    • pp.234-240
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    • 1996
  • A double-stranded RNA binding factor (RBF), characterized as an inhibitor of PKR kinase in our previous study, was evaluated for its RNA binding specificities by RNA gel electrophoretic mobility shift analysis and membrane filter binding assay, RBF displayed affinities for a broad range of RNAs including viral RNAs and synthetic RNAs consiting of stem and loop structures. GC-rich RNA stem helices as short as 11 bp are suggested to represent the minimal binding motif for RBF. RBF binding to all the natural RNAs tested was reversible by poly(I): poly(C) addition, but E. coli 5S RNA was inefficient.

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Development Ability of Bovine Early Embryo Blastomere In Vitro in Embedding Matrix (체외생산 소 초기배 할구세포의 Embedding Matrix에서의 발생능력)

  • 이홍준;서승운;이상호;송해범
    • Journal of Embryo Transfer
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    • v.10 no.2
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    • pp.171-175
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    • 1995
  • This study was performed to establish the condition and the methods for the techniques of insertion the isolated blastomere cells into cytoplasm, in order to research the develop-mental ability of bovine embryo blastomere cells in vitro produced. After 24h in vitro ovary maturation with the ovaries from a slaughter house, in vitro fertilization was performed to the vital sperms which their mobility were decided by percoll gradient method, with 2~8 cell stage embryos, the blastomeres were isolated in $Ca^2$+. $Mg^2$+-free PBS, and following that embedded into agar and alginate solution, respectively. The rates of in vitro develop-ment are as follows ; in agar embedded 11 among 120(9.2%) 1 /2~1 /3 blastomers cleaved and 6 among 93(6.5%) 1 /4~1 /8 blastomeres cleaved. In sodium alginate-embedded 14 among 84(16.7%) 1 /2~1 /3 blastomeres cleaved and 6 among 85(7.1%) 1 /4~1 /8 blastomeres cleaved. In case of Na-alginate, the rate of the cells were better than those of agar. The results suggest that the techniques for embeeding the isolated blastomeres into gel may help cloning of bovine early embryo without nuclear transplantation.

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Evaluation of Electrical Properties of IZO Thin-Film with UV Post-Annealing Treatment Time (IZO 박막 트랜지스터의 UV를 이용한 후열처리 조사 시간에 따른 전기적 특성 평가)

  • Lee, Jae-Yun;Kim, Han-Sang;Kim, Sung-Jin
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.33 no.2
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    • pp.93-98
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    • 2020
  • We investigated the effect of a post-annealing process using ultraviolet (UV) light on the electrical properties of solution-processed InZnO (IZO) thin-film transistors (TFTs). UV light was irradiated on IZO TFTs for different time periods of 0s, 30s, and 90s. We measured transfer and retention stability curves to evaluate the performance of the fabricated TFTs. In addition, we measured height, amplitude, and phase AFM images to analyze changes in the surface and morphology of the devices. AFM measurements were performed by setting the drive amplitude of the cantilever tip to 47.9 mV in tapping mode, then dividing the device surface into 500 nm × 500 nm. In the case of IZO TFT irradiated with UV for 30s, the electron mobility and Ion/Ioff ratio were improved, the threshold voltage was reduced by approximately 2 V, and the subthreshold swing also decreased form 1.34 V/dec to 1.11 V/dec.

Proton Conductivity of Niobium Phosphate Glass Thin Films

  • Kim, Dae Ho;Park, Sung Bum;Park, Yong-il
    • Korean Journal of Materials Research
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    • v.28 no.5
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    • pp.308-314
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    • 2018
  • Among the fuel cell electrolyte candidates in the intermediate temperature range, glass materials show stable physical properties and are also expected to have higher ion conductivity than crystalline materials. In particular, phosphate glass has a high mobility of protons since such a structure maintains a hydrogen bond network that leads to high proton conductivity. Recently, defects like volatilization of phosphorus and destruction of the bonding structure have remarkably improved with introduction of cations, such as Zr4+ and Nb5+, into phosphate. In particular, niobium has proton conductivity on the surface because of higher surface acidity. It can also retain phosphorus content during heat treatment and improve chemical stability by bonding with phosphorus. In this study, we fabricate niobium phosphate glass thin films through sol-gel processing, and we report the chemical stability and electrical properties. The existence of the hydroxyl group in the phosphate is confirmed and found to be preserved at the intermediate temperature region of $150-450^{\circ}C$.

Influence of Fast Neutron Irradiation on the Electrical and Optical Properties of Li Doped ZnSnO Thin Film Transistor (Li 도핑된 ZnSnO 박막 트랜지스터의 전기 및 광학적 특성에 대한 고속 중성자 조사의 영향)

  • Cho, In-Hwan;Kim, Chan-Joong;Jun, Byung-Hyuk
    • Korean Journal of Materials Research
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    • v.30 no.3
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    • pp.117-122
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    • 2020
  • The effects of fast neutron irradiation on the electrical and optical properties of Li (3 at%) doped ZnSnO (ZTO) thin films fabricated using a sol-gel process are investigated. From the results of Li-ZTO TFT characteristics according to change of neutron irradiation time, the saturation mobility is found to increase and threshold voltage values shift to a negative direction from 1,000 s neutron irradiation time. X-ray photoelectron spectroscopy analysis of the O 1s core level shows that the relative area of oxygen vacancies is almost unchanged with different irradiation times. From the results of band alignment, it is confirmed that, due to the increase of electron carrier concentration, the Fermi level (EF) of the sample irradiated for 1,000 s is located at the position closest to the conduction band minimum. The increase in electron concentration is considered by looking at the shallow band edge state under the conduction band edge formed by fast neutron irradiation of more than 1,000 s.

Development of Inverted Organic Photovoltaics with Anion doped ZnO as an Electron Transporting Layer

  • Jeong, Jae Hoon;Hong, Kihyon;Kwon, Se-Hun;Lim, Dong Chan
    • Journal of the Korean institute of surface engineering
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    • v.49 no.6
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    • pp.490-497
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    • 2016
  • In this study, 3-dimensional ripple structured anion (chlorine) doped ZnO thin film are developed, and used as electron transporting layer (ETL) in inverted organic photovoltaics (I-OPVs). Optical and electrical characteristics of ZnO:Cl ETL are investigated depending on the chlorine doping ratio and optimized for high efficient I-OPV. It is found that optimized chlorine doping on ZnO ETL enhances the ability of charge transport by modifying the band edge position and carrier mobility without decreasing the optical transmittance in the visible region, results in improvement of power conversion efficiency of I-OPV. The highest performance of 8.79 % is achieved for I-OPV with ZnO:Cl-x (x=0.5wt%), enhanced ~10% compared to that of ZnO:Cl-x (x=0wt%).