• Title/Summary/Keyword: electro-phoresis

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Bacterial Community Analysis during Composting of Garbage using Denaturing Gradient Gel Electro-phoresis (Denaturing Gradient Gel Electrophoresis를 이용한 음식물 쓰레기 퇴비화 세균 군집 분석)

  • Ryu Hee Wook;Cho Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.33 no.3
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    • pp.226-230
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    • 2005
  • The microbial community during composting of gargage was analyzed using 16S rDNA PCR - DGCE (denaturing gradient gel electrophoresis). Pseudomonas spp. was found throughout the process, and thermophilic Bacillus spp. was dominated at the thermophilic stage. Six thermophilic bacteria were isolated and identified as B. caldoxylolyticus, B. thermoalkalophilus, and B. thermodenitrificans.

Synthesis of $\textrm{N}_{G}$-Mon $o^{14}\textrm{C}$-methyl]-L-arginine ($\textrm{N}_{G}$-Mon $o^{14}\textrm{C}$-methyl]-L-arginine의 합성)

  • 조영봉
    • Environmental Analysis Health and Toxicology
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    • v.1 no.1
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    • pp.77-80
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    • 1986
  • Radiochemical synthesis of $N^{G}$-mono[$^{14}$ C-methyl]-L-arginine is described. The compound was synthesized from radio-active mono[$^{14}$ C]-methylamine as easily and purified by strong cation-exchange resin (NH form) liquid chromatography using a gradient of ammonium hydroxide, and crystallized as flavianate. The free amino acid was successfully prepared by strirring its flavianate and strong anion-exchange resin (OH- form), which could remove the flavianic acid from its salt in water below room temperature. Purity of the compound was tested by thin-layer chromatography, thin-layer electro-phoresis, and scintillation spectrometry.y.

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An Active Monomeric Form of Bovine Milk Xanthine Oxidase

  • Lee, Chu-Hee;Nam, Doo-Hyun;Huh, Keun
    • Archives of Pharmacal Research
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    • v.17 no.5
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    • pp.318-322
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    • 1994
  • Upon gel filtration, the commercial bovine milk xanthine oxidase preparation was fractionated into two preparations showing enzyme activity. Native polyacrylamide gel electrophoresis showed that one was in a dimeric form and the other was a monomer having molecular weight of 150 kDa. It was also found that this commercial enzyme existed mostly in an active monomeric form without loss of enzyme activity. The rabbit antisera produced against two enzyme preparations cross-reacted well each other. In SDS-polyacrylamide gtel electro-phoresis, however, both enzyme preparations yielded two smaller protein bands below 150 kDa, which appeared to bind with both antisera with high affinity but not to retain enzyme activity. It implies that bovine milk xanthine oxidase can lose its activity when monomeric subunit is further degraded.

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PCR Analysis of Four Length-Polmorphic Loci in Korea Population for Genotyping

  • Ryu, Jae-Song;Koo, Yoon-Mo;So, Jae-Seong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.3
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    • pp.169-173
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    • 2000
  • On human chromoscomes, a short sequence of DNA is known to repeat a number of times. These are called variable number of tandem repeat (VNTR) or short tandem respeat (STR) which has a short core. VNTR and STR are used in the filed of forensic science, evolution, and anthropology. In this work, we examined allele frequencies of one VNTR (YNZ22) and three STRs (NeuR, D21S11, Humth01) in a korean population sample by polymerase chain reaction (RCP) followed by high-resolution polyacrylamide gel electro-phoresis (PAGE) with silver stain. Subsequently, the polymorphism information content (PIC) was calculated : the hifhest PIC was observed in the NeuR locus (0.95680) and lowest in the Humth01 locus (0.75809).

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Identification of the Chicken Meat Allergens (닭고기 중 알레르기 유발성분의 동정)

  • 조은득;김동섭;정기화
    • Biomolecules & Therapeutics
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    • v.9 no.1
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    • pp.7-14
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    • 2001
  • The chicken meat has been reported as one of the food causing allergic reactions predominantly to Korean. At present, several in vitro tests for immunoglobulinG (IgG)-mediated as well as IgE-mediated food allergy are available. 13 clinically chicken meat-allergic patients were investigated together with 4control subjects for identification of chicken meat-specific reactivity by ELISA. Also, protein profile and IgE, IgGtotal and IgG4-reacting allergens were detected by means of sodium dodecyl sulfate-polyacrylamide gel electro-phoresis (SDS-PAGE)and immunoblotting. Chicken meat extracts were prepared as raw, heated, heat and simulated gastric fluid (SGF) treated samples to characterize the stability of allergen to physicochemical treatment. SDS-PAGE revealed 9~200 kDa bands. And in immunoblotting 7 sera were identified most major bands between 10 and 78 kDa. In case of IgE, six proteins (17, 26, 35, 40, 78 kDa) were predominant in heat-treated extract, and the one (35 kDa) was present in SGF-treated preparations. In case of IgG$_{total}$ and IgG4, most of them showed a patters simmilar to IgE. There were significant differences (P<0.05) in IgE, IgG$_{total}$ , IgG4 Abs to chicken meat between the allergic and control subjects in ELISA. In addition, the concentration of IgG4Abs in the challenge-positive subjects was significantly higher than that of control subjects. It is considered that the specific IgE response to chicken meat was rarely prevalent to Koreans. However, the specific IgG4 response play an important role in the development of allergic symptoms.

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Fabrication of electro phoresis microchips and effects of channel surface properties (마이크로 전기영동 소자의 제작과 유로 면 특성에 따른 전기삼투 및 전기영동 효과)

  • Kim, Min-Su;Cho, Seung-Il;Lee, Kook-Nyung;Kim, Yong-Kweon
    • Proceedings of the KIEE Conference
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    • 2003.10a
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    • pp.286-289
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    • 2003
  • We investigated the influence of the properties of substrate material on the separation efficiency in microchip electrophoresis. We fabricated the various microchips and studied separation efficiency in microchannels composed of a single material such as quartz, glass, polydimethylsiloxane (PDMS), and polymethylmetha crylate (PMMA), as well as hybrid micro channels composed of different materials. New fabrication process for glass chip was suggested and some treatment is added to improve fabrication process in other chip. Separation efficiency was compared by measuring migration times and bandwidths of EOF and analytes in each microchip. The efficiency is the function of migration time, which is affected by the electroosmotic flow (EOF), and bandwidth of an analyte. EOF is highly dependent upon the characteristics of a microchannel wall surface. Migration time was more reproducible in silica chips than that of PDMS chip and more band broadening was observed in the microchip composed of hybrid material due to non-uniformity of surface charge density at the walls of the channel.

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Enantioselective Pharmacokinetics of Carvedilol in Human Volunteers

  • Phuong, Nuyen-Thi;Lee, Beam-Jin;Choi, Jung-Kap;Kang, Jong-Seong;Kwon, Kwang-il
    • Archives of Pharmacal Research
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    • v.27 no.9
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    • pp.973-977
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    • 2004
  • Carvedilol is administered as a racemic mixture of the R(+)- and S(-)-enantiomers, although they exhibit different pharmacological effects. To investigate the stereoselective pharmacoki-netics, the enantiomeric separation of carvedilol in human plasma was undertaken using capil-lary electrophoresis (CE). Resolution of the enantiomers was achieved using 2-hydoxypropyl-$\beta$-cyclodextrin as the chiral selector. Phosphate buffer (50 mM, pH 4.0) containing 10 mM of 2-hydoxypropropyl-$\beta$-cyclodextrin was used as electrolytic buffer. Achiral separation was carried out with the same electrolytic buffer without chiral selector. Following a single oral administra-tion of 25-mg carvedilol to 11 healthy, male volunteers, stereoselective pharmacokinetic analy-sis was undertaken. The maximum plasma concentrations ( $C_{max}$) were 48.9 and 21.6 ng/mL for (R)-carvedilol and (S)-carvedilol, respectively, determined by the chiral method. The profiles of the plasma concentration of (RS)-carvedilol showed $C_{max}$ of 71.5, 72.2, and 73.5 ng/mL, as determined by the CE, HPLC/FD methods and calculations from the data of the chiral method, respectively.y.y.

Isolation, Purification and Characterization of Phytase from Asperfillus sp. (Aspergillus속 균주가 생산하는 Phytase의 분리 정제 및 특성)

  • 천성숙;조영제;차원섭;이희덕;이선호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.1
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    • pp.38-45
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    • 1998
  • To extract insoluble proteins and to improve funtional properties of abolished proteins, an phytase producing Aspergillus sp. SM-15 was isolated from soil. The enzyme was purified and its enzymological characteristics were investigated. Phytase production reached to maximum when the wheat bran medium containing 1% mannose, 1% yeast extract, 1% (NH4)2HPO4 and 0.2% calcium chloride was cultured for 4 days. Phytase was purified 17.1 fold and specific activity was 244.32unit/mg by a sequencial process of ammonium sulfate fraction, ion exchange chromatography and gel filtrations Pruified enzyme was confirmed as a single band by the polyacrylamide gel electro-phoresis. The molecular weight of phytase was estimated to be 46,000. The optimum pH and temperature for the phytase activity were 5.5 and 5$0^{\circ}C$. The enzyme is stable in pH 4.5~5.5, 6$0^{\circ}C$. The activity of purified enzyme was inhibited by Hg2+ whereas activited by Pb2+ and Fe2+. The activity of phytase was inhibited by the treatment with iodine. The result indicate the possible involvement of histidine at active site. Km and Vmax of the puridied phytase were 37.037mM/L and 159.87umol/min, respectively.

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Differential protein expression in avian liver in response to invasion by Salmonella gallinarum

  • Lee, Gang-Deog;Cho, In-Hee;So, Hyun-Kyung;Koo, Yong-bum;Lee, Jun-heon;Choi, Kang-Duk
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2004.11a
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    • pp.37-38
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    • 2004
  • Salmonella gallinarum is a pathogen that is capable of causing disease in Korean native chicken. Although Salmonella gallinarum is important world-wide pathogens of poultry, little is understood of the mechanisms of pathogenesis of Salmonella gallinarum in the chicken. This study was to investigate chicken liver proteins affected by infection of Salmonella gallinarum in Korean native chicken. The differentially expressed proteins of chicken livers were identified by using 2-dimensional electro- phoresis (2D-E) and mass spectrometry (MS). We detected more than 300 protein spots on silver stained 2D gels using pH 3∼10 gradients. Three differentially expressed protein spots were analyzed by MALDI-TOF MS and MS/MS. The obtained MS and MS/MS data were searched against a protein database using the Mascot search engine. Further researches on the identified proteins can give valuable information of mechanism of pathogenesis in chicken.

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Characterization and Partial Nucleotide Sequence of Potato Virus X Isolated from Potato in Korea

  • Jung, Hyo-Won;Yun, Wan-Soo;Seo, Hyo-Won;Hahm, Young-Il;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.16 no.2
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    • pp.110-117
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    • 2000
  • Potato virus X (PVX-KO) showing mild mosaic and stunting symptoms on potato (Solanum tuberosum) in Kangwon area has been isolated and characterized. EM observation of the purified virus particles showed flexuous rod shape of about 520 nm in length. The coat protein (CP) of the virus had a molecular weight of 31 kDa in SDS-PAGE analysis, and the viral RNA was approximately 6.4 kb in size in denatured agarose gel electro-phoresis. In gel-immunodiffusion tests, it reacted strongly with an antiserum to common PVX from BIOREABAAG (USA). A rabbit antiserum was produced using purified virus and used for routine PVX detection by ELISA. Cultivated potatoes in Kangwon and other areas were frequently infected with PVX-KO. Both Datura stramonium and Nicotiana tabaccum cultivars developed necrotic local lesions 5 days after inoculation, and systemic mosaic symptoms with vein clearing 2 weeks after inoculation. All the features agree with the description of other PVX strains. To confirm and determine PVX strains, reverse transcription-polymerase chain reaction experiment was conducted using specific primers for viral CP. Amplified DNA fragments were cloned and sequenced. Results showed nucleotide sequence homologies of about 88 to 99% to other PVX strains. Based on CP amino acid sequence deduced from nucleotide sequences and host range studies PVX-KO is considered a member of the type X subgroup of PVX.

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