• Title/Summary/Keyword: 2D gel electrophoresis

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Molecular-epidemiologic study on outbreak of colonization by extended spectrum β-lactamase producing Klebsiella pneumoniae in neonatal intensive care unit (신생아 중환자실에서 extended spectrum β-lactamase를 생성하는 Klebsiella pneumoniae 집단 보균 발생의 분자 역학적 조사 및 추적관찰)

  • Jun, Nu-Lee;Kim, Mi-Na;Jeong, Jae-Sim;Kim, Yang-Soo;Kim, Ellen Ai-Rhan;Kim, Ki-Soo;Pi, Soo-Young
    • Clinical and Experimental Pediatrics
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    • v.49 no.2
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    • pp.150-156
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    • 2006
  • Purpose : The aims of this study included assessment of molecular-epidemiologic features during an outbreak of colonization of extended spectrum ${\beta}$-lactamase producing Klebsiella pneumoniae(ESBL-KPN) and re-evaluation of their colonized status one year later. Methods : Rectal swab cultures for ESBL-KPN from all hospitalized infants and newly admitted infants were obtained during the outbreak of colonization from July to December, 2000. The pattern of XbaI-digested chromosomal DNA of isolates were analyzed by pulsed-field gel electrophoresis. Weekly rectal swab cultures were obtained during the outbreak until patients were either discharged or decolonized. Patients discharged after being colonized had follow up stool cultures a year later. Results : A total of 80 patients(28.5 percent) were colonized. Of those, 53 whose pulsed-field gel electrophoresis(PFGE) was possible only once, were ESBL-KPN grouped into six cluster clones and 10 single clones : 28 patients(52.8 percent) were colonized with type A, the most common clone, followed by type B in 11 patients(20.8 percent). Of those 12 patients in whom serial PFGE was done more than twice, type A was predominant. Narrowed-down in strains occurred from types A, B, C, D and three single clones at initiation of the study into types A and type B after three months of strict infection control. Among 75 patients(93.7 percent) who were sent home after being colonized, 30 patients were re-called for stool cultures a year later : All of them were decolonized. Conclusion : This study demonstrates the importance of infection control as the diversity of ESBL-KPN strains could be narrowed into fewer strains. Colonization of ESBL-KPN could be reversed upon return to the community.

Differential Proteome Analysis of Breast and Thigh Muscles between Korean Native Chickens and Commercial Broilers

  • Liu, Xian De;Jayasena, Dinesh D.;Jung, Yeon-Kuk;Jung, Samooel;Kang, Bo-Seok;Heo, Kang-Nyeong;Lee, Jun-Heon;Jo, Cheo-Run
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.6
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    • pp.895-902
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    • 2012
  • The Korean native chickens (Woorimotdak$^{TM}$, KNC) and commercial broilers (Ross, CB) show obvious differences in meat flavor after cooking. To understand the contribution of protein and peptide for meat flavor, 2-dimensional (2-D) gel electrophoresis and matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry was performed. A total of 16 protein spots were differentially expressed in the breast and thigh meat between the two breeds. A total of seven protein spots were represented by different levels between KNC and CB for breast meat. Among them three protein spots (TU39149, TU40162 and TU39598) showed increases in their expressions in KNC while other four protein spots (BU40125, BU40119, BU40029 and BU39904) showed increases in CB. All nine protein spots that were represented by different levels between KNC and CB for thigh meat showed increases in their expression in KNC. Phosphoglucomutase 1 (PGM 1), myosin heavy chain (MyHC), heat shock protein B1 (HSP27), cytochrome c reductase (Enzyme Q), Glyoxylase 1, DNA methyltransferase 3B (DNA MTase 3) were identified as the main protein spots by MALDI-TOF mass spectrometry. These results can provide valuable basic information for understanding the molecular mechanism responsible for breed specific differences in meat quality, especially the meat flavour.

YlaC is an Extracytoplasmic Function (ECF) Sigma Factor Contributing to Hydrogen Peroxide Resistance in Bacillus subtilis

  • Ryu Han-Bong;Shin In-Ji;Yim Hyung-Soon;Kang Sa-Ouk
    • Journal of Microbiology
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    • v.44 no.2
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    • pp.206-216
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    • 2006
  • In this study, we have attempted to characterize the functions of YlaC and YlaD encoded by ylaC and ylaD genes in Bacillus subtilis. The GUS reporter gene, driven by the yla operon promoter, was expressed primarily during the late exponential and early stationary phase, and its expression increased as the result of hydrogen peroxide treatment. Northern and Western blot analyses revealed that the level of ylaC transcripts and YlaC increased as the result of challenge with hydrogen peroxide. A YlaC-overexpressing strain evidenced hydrogen peroxide resistance and a three-fold higher peroxidase activity as compared with a deletion mutant. YlaC-overexpressing and YlaD-disrupted strains evidenced higher sporulation rates than were observed in the YlaC-disrupted and YlaD-overexpressing strains. Analyses of the results of native polyacrylamide gel electrophoresis of recombinant YlaC and YlaD indicated that interaction between YlaC and YlaD was regulated by the redox state of YlaD in vitro. Collectively, the results of this study appear to suggest that YlaC regulated by the YlaD redox state, contribute to oxidative stress resistance in B. subtilis.

The Changes of Casein of Cheese Base Treated with Enzyme during Ripening (Enzyme의 처리에 의한 Cheese Base의 숙성중 Casein의 변화)

  • Lee, Gang-Ik;Cha, Gwang-Jong;Yu, Je-Hyeon
    • Journal of Dairy Science and Biotechnology
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    • v.14 no.2
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    • pp.135-146
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    • 1996
  • This experiment was carried out to investigate the changes of casein of cheese base treated with substitute enzyme during ripening. The cheese base without enzyme treatment(control, D)and cheese base treated with only calf rennet(A), cheese base treated with mixed enzyme(calf rennet :porcine pepsin 1:1, B), cheese base treated with only porcine pepsin(C) were manufactured. The changes of casein were analyzed by means of HPLC and electrophoresis as experimental parameters during ripening. Gel filtration(HPLC) of casein by Superose 12 column in Cheddar cheese showed 5 fractions immediately after manufacturing and 8 fractions after six months ripening. Though D showed no difference in number of fraction(4 fraction) during 8 weeks ripening, A, B, C have represented the change of fraction number 4 to 5, 4 to 7, 4 to 8, respectively. As the mixing ratio of porcine pepsin increased, higher degradability of casein appeared. After 8 weeks ripening, electrophoresis of casein in cheese base showed three bands as an ${\alpha}$$_{s1}$casein from A and five bands from B, C. In case of D one major band and two minor bands were appeared as an ${\alpha}$$_{s1}$-casein. As the additional level of porcine pepsin increased the concentration of ${\beta}$-casein band decreased. however, that of ${\gamma}_1$ ${\gamma}_2$-casein band increased and para-${\kappa}$-casein band appeared from A, B, C, except D.

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Electrophoretic Patterns of Hemolymph Protein of Healthy and Nuclear-Polyhedrosis Virus Infected Silkworm, Bombyx mori. (건강잠 및 핵다각체 이병잠의 체액단백질의 전기영동상)

  • 김낙정;강석권;김정일
    • Journal of Sericultural and Entomological Science
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    • v.13 no.2
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    • pp.141-143
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    • 1971
  • By means of thin-layer electrophoresis in agarose gel, hemolymph protein of healthy silkworm larvae and of the nuclear polygedrosis virus infected larvae were studied. 1. In the 4th instar, 4 fractions moving toward anode were separated. Dye-binding Capacity of the fraction was increased according to the stage. 2. After 5th day in the 5th instar, 7 fractions moving toward anode were separated, and one fraction toward cathode was separated. 3. On the first day in the 5th instar, 5 fractions were separated, and on the 4th day of the same instar 5 fractions were separated. 4. As for the hemolymph protein fractions of the polyhedrosis virus infected larvae, on the 6th and 7th day, three fractions(D.E.F) were inclined to increase, whereas on the 8th day 4 fractions(A.B.D.E) were disappeared but F fraction was inclined to decrease.

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Isolation and Characterization of Lectin in Soybean(Glycine max L.) (대두(Glycine max L.)의 렉틴 분리 및 특성)

  • 박원목;이용세;박상호;김성환;윤경은
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.34 no.2
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    • pp.120-126
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    • 1989
  • This experiment was carried out to investigate the lectin of soybean (Glycine max L.) seed. Purification was done by 50-80% ammonium sulfate precipitation, CM-cellulose and Sephadex G-100 column. The purity was ascertained by electrophoresis. The molecular weight of purified lectin was estimated as 132,000. It was composed of three subunits which molecular weight was 45,000. The lectin was identified as glycoprotein by Schiff's reagent staining and Dubois method. The lectin agglutinated erythrocytes of rabbit and human. The amounts of the lectin to agglutinate human erythrocytes differed among the blood types: The blood type A required the least amount, the next was B, O, and AB in order. The agglutination was specifically inhibited by 5${\mu}$g/ml of N -acetyl.-D-galactoseamine and 200${\mu}$g/ml of D-galactose. Other tested sugars could not inhibit the agglutination of the erythrocytes by the lectin.

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Chrysanthemum stunt viroid in Dendranthema grandiflorum

  • Chung, Bong-Nam;Park, Gug-Seoun;Kim, Hyun-Ran;Kim, Jeong-Soo
    • The Plant Pathology Journal
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    • v.17 no.4
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    • pp.194-200
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    • 2001
  • Chrysanthemum stunt viroid (CSVd) ws identified in chrysanthemum cv. Chunkwang showing symptoms of stunt with leaf distortion (K1) and stunt with chlorosis of leaves (K2) collected from the main cultivation area of Masan, Kyongnam province in Korea. The specific RNAs related with the diseased chrysanthemums were detected. Full-length 354 bp CSVd cDNAs were amplified from infected tissue by reverse transcription and polymerase chain reaction using a pair of primers specific for CSVd sequence. The amplified cDNA products were analyzed by agarose gel electrophoresis and the specific cDNAs were cloned. Nucleotide sequences of the two CSVd isolates K1 and K2 varied. Phylogenetic analysis of the nucleotide sequences of CSVd isolates indicated that K1 was closely related with J2 and Am 2 isolates. K1 and K2 were transmitted by grafting to Dendranthema grandiflorum cv. Mistletoe, Gynura aurantiaca, and Lycopersicon esculentum cv. Rutgers. This is the first report of CSVd in D. grandiflorum in Korea.

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Proteome Analysis of Bovine Longissimus dorsi Muscle Associated with the Marbling Score

  • Shen, Y.N.;Kim, S.H.;Yoon, D.H.;Lee, H.G.;Kang, H.S.;Seo, K.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.8
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    • pp.1083-1088
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    • 2012
  • The breeding value of marbling score in skeletal muscle is an important factor for evaluating beef quality. In the present study, we investigated proteins associated with the breeding value of the marbling score for bovine sirloin to select potential biomarkers to improve meat quality through comparative proteomic analysis. Proteins isolated from muscle were separated by two-dimensional gel electrophoresis. After analyzing images of the stained gel, seven protein spots for the high marbling score group were identified corresponding to changes in expression that were at least two-fold compared to the low marbling score group. Four spots with increased intensities in the high marbling score group were identified as phosphoglycerate kinase 1, triosephophate isomerase, acidic ribosomal phosphoprotein PO, and capping protein (actin filament) Z-line alpha 2. Spots with decreased intensities in the high marbling score group compared to the low score group were identified as 14-3-3 epsilon, carbonic anhydrase II, and myosin light chain 1. Expression of myosin light chain 1 and carbonic anhydrase 2 was confirmed by Western blotting. Taken together, these data could help improve the economic performance of cattle and provide useful information about the underlying the function of bovine skeletal muscle.

Isolation of Plasmids from the Moderately Halophilic Bacteria (Moderate 호염성 세균의 Plasmid 유전자 분리)

  • HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.18 no.6
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    • pp.557-562
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    • 1985
  • Moderately halophilic bacteria were collected from solar salt with Larsen medium containing $10\%$ NaCl. A total of 56 strains were isolated and tested for the presence of plasmid DNA by agarose gel electrophoresis. Twelve isolates ($21\%$) carried at least one kind of plasmid. Six different isolates among them were selected to study the molecular weight of plasmids and the morphological and physiological characters. Vibrio sp. 14, Alcaligenes sp. 63, Pseudomonas sp. 11, Flavobacterium sp. 38, Bacillus sp. 16, and Alcaligenes sp. 52 carried at least one plasmid of about 7.2 kbp, 6.4 kbp, 6.85 kbp, 8.5 kbp, 8.75 kbp, and 6.8 kbp respectively.

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Monitoring of Microbial Diversity and Activity During Bioremediation of Crude Oil-Contaminated Soil with Different Treatments

  • Baek, Kyung-Hwa;Yoon, Byung-Dae;Kim, Byung-Hyuk;Cho, Dae-Hyun;Lee, In-Sook;Oh, Hee-Mock;Kim, Hee-Sik
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.67-73
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    • 2007
  • The present study compared the microbial diversity and activity during the application of various bioremediation processes to crude oil-contaminated soil. Five different treatments, including natural attenuation (NA), biostimulation (BS), biosurfactant addition (BE), bioaugmentation (BA), and a combined treatment (CT) of biostimulation, biosurfactant addition, and bioaugmentation, were used to analyze the degradation rate and microbial communities. After 120 days, the level of remaining hydrocarbons after all the treatments was similar, however, the highest rate (k) of total petroleum hydrocarbon (TPH) degradation was observed with the CT treatment (P<0.05). The total bacterial counts increased during the first 2 weeks with all the treatments, and then remained stable. The bacterial communities and alkane monooxygenase gene fragment, alkB, were compared by denaturing gradient gel electrophoresis (DGGE). The DGGE analyses of the BA and CT treatments, which included Nocardia sp. H17-1, revealed a simple dominant population structure, compared with the other treatments. The Shannon-Weaver diversity index (H') and Simpson dominance index (D), calculated from the DGGE profiles using 16S rDNA, showed considerable qualitative differences in the community structure before and after the bioremediation treatment as well as between treatment conditions.