• Title/Summary/Keyword: E-Isolation

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Translation Initiation Factor IF1-Dependent Stimulation of 30 S Preinitiation Complex Formation: Rapid Isolation and fMEt-tRNA Binging Activity of IF1

  • CHOIK, SANG-YUN;HYUN-JUNG KIM;JUNG-IK YANG;HYO-IL CHANG
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.986-993
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    • 2001
  • Translation Initiation in prokaryotes involves the formation of a 30 S preinitiation complex, in which translation initiation factors play a role in the stimulation of fMet-tRNA (fMet) binding. However, the specific function and precise mechanism of initiation factor IF1 are still unclear. One a functionally active factor with a high purity. In the present study a large quantity of active IF was rapidly purified, obtained by the overexpression of the infA gene, and then used for a functional study. The induction of infA did not appreciably affect the growth rate of the protease-deficient strain E. coli AR68 harboring the IF1 overproducing plasmid. The level of IF1 obtained was approximately $1-2\%$ of the total cell protein, which enabled the yield of highly purified IF1 (>$98\%$ pure) to be increased to 0.15 mg of IF1/g of cells. The IF1 was isolated within one day by the centrifugatioin of the ribosomal washed fraction, by ammonium sulfate fractionation, chromatography on batch of phosphocellulose, and FPLC Mono S. The overexpressed IF1 was found to be comparable to the factor isolated from normal cells, as determined by migration in NEPHGE/SDS 2-D gels. For binding of fMet-tRNA(fMet) to the 30 S ribosomal subunitis, relatively high levels of binding were obtained when IF2 was present. The addition of IF1 up to 110 pmol proportionally stimulated the binding to a variable extent. This IF1-dependent stimulation of the 30 S preinitiation complex formation demonstrated that IF1 would appear to be exclusively essential for promoting the initiation phase of protein synthesis.

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Isolation and Characterization of Chlorothalonil-dissipating Bacteria from Soil. (토양으로부터 Chlorothalonil 전환 미생물의 분리 및 특성)

  • 이수현;신재호;최준호;박종우;김장억;이인구
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.96-100
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    • 2004
  • Chlorothalonil is a wide-spectrum fungicide that is widely used in the world. Chlorothalonil is known as a potential toxic pollutant due to its high application rate, persistence, and toxicity to humans and other species. With the Increase of necessity of bioremediation, this study was conducted to isolate the chlorothalonil dissipation bacteria from soil. Soil samples were collected from 184 sites of farmland and wastewater disposal soil.661 strains resistant to chlorothalonil were isolated by dilution method from chlorothalonil-containing enrichment culture. After incubating at $30^{\circ}C$ in 1/10 LB media containing 10 ppm of chlorothalonil for a week, dissipation ability of chlorothalonil was investigated by HPLC. Finally, a strain SH35B, capable of dissipating chlorothalonil efficiently, was selected. The strain SH35B was identified as Ochrobactrum sp. Ten ppm of chlorothalonil In 1/10 LB media were completely dissipated by the growth of Ochrobactrum sp. SH35B for 30 h at $30^{\circ}C$. In the isolated strain, the content of glutathione and the activity of glutathione S-transferase were supposed to be ones of the Important factors for chlorothalonil dissipation and were higher than those of control strains, Escherichia coli and Bacillus subtilis.

Establishing optimal gap size for precast beam bridges with a buffer-gap-elastomeric bearings system

  • Farag, Mousa M.N.;Mehanny, Sameh S.F.;Bakhoum, Mourad M.
    • Earthquakes and Structures
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    • v.9 no.1
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    • pp.195-219
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    • 2015
  • A partial (hybrid) seismic isolation scheme for precast girder bridges in the form of a "buffer-gap-elastomeric bearings" system has been endorsed in the literature as an efficient seismic design system. However, no guides exist to detail an optimal gap size for different configurations. A numerical study is established herein for different scenarios according to Euro code seismic requirements in order to develop guidelines for the selection of optimal buffer-gap arrangements for various design cases. Various schemes are hence designed for ductile and limited ductility behavior of the bridge piers for different seismic demand levels. Seven real ground records are selected to perform incremental dynamic analysis of the bridges up to failure. Bridges with typical short and high piers are studied; and different values of initial gaps at piers are also investigated varying from a zero gap (i.e., fully locked) condition up to an initial gap at piers that is three quarters the gap left at abutments. Among the main conclusions is that the as-built initial gaps at piers (and especially large gap sizes that are ${\geq}1/2$ as-built gaps at abutments) do not practically reduce the seismic design demand and do not affect the reserve capacity of the bridge against failure for bridges featuring long piers, especially when these bridges are designed a priori for ductile behavior. To the contrary, the "buffer-gap-elastomeric bearings" system is more effective for the bridge schemes with short piers having a large difference between the stiffness of the bearings and that of their supporting (much stiffer) squat piers, particularly for designs with limited ductility. Such effectiveness is even amplified for the case of larger initial as-built gap sizes at piers.

Isolation and Identification of FSHβ and LHβ Subunits in the Blacktip Grouper Epinephelus fasciatus (홍바리(Epinephelus fasciatus) FSHβ와 LHβ subunit의 분리 및 동정)

  • Park, Yong-Ju;Kang, Hyeong-Cheol;Lee, Chi-Hoon;Song, Young-Bo;Baek, Hea-Ja;Kim, Hyung-Bae;Soyano, Kiyoshi;Lee, Young-Don
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.45 no.1
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    • pp.17-24
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    • 2012
  • We cloned and sequenced the cDNA encoding the $FSH{\beta}$ and $LH{\beta}$ subunits from the pituitary of the blacktip grouper Epinephelus fasciatus, which regulate vitellogenesis and maturation in vertebrates, to achieve stable and healthy gametes. The full-length cDNAs of $FSH{\beta}$ and $LH{\beta}$ were 571 bp and 617 bp, encoding 120 amino acid (aa) and 147 aa proteins, respectively. The deduced amino acid sequences of $FSH{\beta}$ and $LH{\beta}$ were highly homologous (68-97%) to those of other Perciformes; E. bruneus, Dicentrarchus labrax, Thunnus thynnus, and Pseudolabrus sieboldi. Phylogenetic analysis showed that the deduced $FSH{\beta}$ and $LH{\beta}$ amino acid sequences were categorized as a distinct subunit in the $GTH{\beta}$ family, and are closely related to the teleostei $FSH{\beta}$ and $LH{\beta}$, respectively. $FSH{\beta}$ and $LH{\beta}$ mRNA exhibited high abundance in the pituitary gland and low in other brain areas, but were not present in peripheral tissues, as determined by RT-PCR.

Cloning of Mouse AQP-CD Gene

  • Jung, Jin-Sup;Kim, Joo-In;Oh, Sae-Ok;Park, Mi-Young;Bae, Hae-Rhan;Lee, Sang-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.2
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    • pp.195-200
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    • 1997
  • Water transport in highly-permeable membranes is facilitated by some specialized pathways, which are called aquaporins (AQP). AQP1 (AQP-CHIP) is the first recognized aquaporin identified from red cells and renal proximal tubules. Up until now 4 other aquaporin homologs have been reported. Each aquaporin has its unique tissue distribution and regulatory mechanims. To elucidate molecular mechanisms for their transcription regulation and tissue-specific expression isolation of aquaporin genes is required. To clone promoters of the AQP family mouse genomic library was screened by the 1st exon-specific probe of AQP4, and 5 different plaques were positively hybridized. Phage DNAs were purified and characterized by restriction mapping and sequencing. One of them is the mouse AQP-CD gene. The gene was consisted of 4 exons and the exon-intron boundaries of mouse AQP-CD gene were identified at identical positions in other related genes. The 5'-flanking region of AQP-CD gene contains one classic TATA box, a GATA consensus sequence, an E-box and a cyclic AMP-responsive element. The cloning of the mouse AQP-CD gene, of which product is expressed in the collecting duct and is responsible for antidiuresis by vasopressin, will contribute to understand the molecular mechanisms of tissue-specific expression and regulation of AQP-CD gene under various conditions.

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'Survey on Bacteriological Contamination of Moving Tavern in Seoul Area' ('노상주점의 위생상태에 관한 미생물학적 조사')

  • Yu Byong Tai
    • Journal of environmental and Sanitary engineering
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    • v.1 no.1 s.1
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    • pp.59-67
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    • 1986
  • This sanitary survey was carried out to investigate the bacteriological contamination of cooking utensils and foods of moving tavern in eight sample sites of Seoul area. The results of survey were as follows: 1. The counts by means of total bacteria in cooking utensils and food samples by standard plate count method were as follow: $5.6\times10^5$ per gm in dishcloth, $3.1\times10^6$ per ml in dishwater. In food samples, $5.4\times10^5$ per gm in meat was higher than other samples. 2. The average counts total coliform and fecal coliform in samples by MPN method were as follow: $3.4\times10^4$ MPN per 100ml, and $1.3\times10^2$ MPN per 100ml in chopping board, $6.1\times10^4$MPN per gm and $1.0\times10^2$ MPN per gm in dishcloth, $1.8\times10^5$ MPN per 100ml and $6.1\times10^2$ MPN per 100ml in dishwater. In food samples, $3.1\times10^4$MPN per gm and $2.0\times10^2$ MPN per gm in meat was higher than other samples. 3. The counts by means of Pseudomonas in samples by MPN method were as follow: $2.8\times10^3$ MPN per 100ml in chopping board, $4.7\times10^3$ MPN per gm in dishcloth $5.6\times10^3$ MPN per 100ml in dishwater. In food samples, $2.4\times10^3$ MPN per gm in shellfish was higher than other samples. 4. Isolation cases of Food poisoning organisms from samples were as follow: Staphylococci was detected 9 cases $(17.6\%)$ in chopping board, 7 cases $(13.6\%)$ in dishcloth. In food samples, 9 cases $(25.7\%)$ in meat, 1 case $(4\%)$ in fish samples. Salmonella was detected 2 cases $(3.9\%)$ in dishwater, 1 case in meat samples.

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Multiwalled Carbon Nanotubes Functionalized with PS via Emulsion Polymerization

  • Park, In-Cheol;Park, Min;Kim, Jun-Kyung;Lee, Hyun-Jung;Lee, Moo-Sung
    • Macromolecular Research
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    • v.15 no.6
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    • pp.498-505
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    • 2007
  • This study demonstrated the in-situ functionalization with polymers of multi-walled carbon nanotubes (MWNTs) via emulsion polymerization. Polystyrene-functionalized MWNTs were prepared in an aqueous solution containing styrene monomer, non-ionic surfactant and a cationic coupling agent ([2-(methacryloyloxy)ethyl]trime-thylammonium chloride (MATMAC)). This process produced an interesting morphology in which the MWNTs, consisting of bead-string shapes or MWNTs embedded in the beads, when polymer beads were sufficiently large, produced nanohybrid material. This morphology was attributed to the interaction between the cationic coupling agent and the nanotube surface which induced polymerization within the hemimicellar or hemicylindrical structures of surfactant micelles on the surface of the nanotubes. In a solution containing MATMAC alone without surfactant, carbon nanotubes (CNTs) were not well-dispersed, and in a solution containing only surfactant without MATMAC, polymeric beads were synthesized in isolation from CNTs and continued to exist separately. The incorporation of MATMAC and surfactant together enabled large amounts of CNTs (> 0.05 wt%) to be well-dispersed in water and very effectively encapsulated by polymer chains. This method could be applied to other well-dispersed CNT solutions containing amphiphilic molecules, regardless of the type (i.e., anionic, cationic or nonionic). In this way, the solubility and dispersion of nanotubes could be increased in a solvent or polymer matrix. By enhancing the interfacial adhesion, this method might also contribute to the improved dispersion of nanotubes in a polymer matrix and thus the creation of superior polymer nanocomposites.

Isolation of antibiotic resistant Lactobacillus spp from fermented milk and starters, and detection of transmissible R plasmids (유산균 발효유 및 starter에서 항생제내성 유산균 분리 및 전달성 R plasmids 분포조사)

  • Baek Kui-Jeong;Seo Heyng-Seok;Roh Young-Sun;Yang Hae-Dong;Hur Boo-Hong;Seo Lee-Won;Joung Dong-Suk;Song Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.29 no.3
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    • pp.277-285
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    • 2006
  • Plasmids are covalently closed circular molecules of DNA that are stably inherited and replicate somewhat independently of the bacterial chromosome. Genes carried on plasmids can mediate a wide variety of important functions, including antibiotics (R plasmids) and heavy metals resistance, toxins production, cell penetration, iron chelation, complement resistance, and metabolic characteristics such as sucrose and lactose fermentation. Fifty strains of lactobacilli were isolated from 26 staters and 29 fermented milk products. They were classified 27 strains as Lactobacillus paracasei subsp paracasei, 11 stains as Lactococcus lactis subsp cremoris, 6 strains as L delbrueckii subsp lactis, 4 strains as L acidophius, and 2 strains as L delbrueckii subsp bulgaricus. All of these strains were examined for drug resistance and transferability of R plasmids. All of the isolates were sensitive to Am, C, CF, E, NB, P, T, and Te. But resistant to SXT 94% (47 strains), K 66% (33 strains), S 56% (28 strains), ENR 50% (25 strains), NOR 38% (19 strains) CIP 38% (19 strains), GM 16% (8 strains), and N 14% (7 strains), in order. And 32 different resistant patterns were found. The most frequently encountered patterns were CIP-ENR-K-NOR-S-SXT (5 strains). In vitro R plasmids transfer experiment, 57 antibiotic resistant strains which were not transfer to the recipient 2 Escherichia coli strains by conjugation, These results indicate that Lactobacillus in internal trade market' stater recognize R factor but transmissible R plasmid is not existed.

Characterization and isolation of shiga toxin-producing Escherichia coli from Bovine feces and Carcass (소의 분변과 도체에서 shiga toxin-producing Escherichia coli의 분리와 특성)

  • Chae, Hee-Sun;Kim, Neung-Hee;Han, Hye-Jin;Son, Hong-Rak;Kim, Chang-Ki;Kim, Sun-Heung;Lee, Jung-Hark;Kim, Jong-Taek
    • Korean Journal of Veterinary Service
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    • v.32 no.3
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    • pp.241-249
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    • 2009
  • Shiga toxin (Stx)-producing Escherichia coli (STEC) strains can cause broad spectrum of human disease, including diarrhea, hemorrhagic colitis, and the life-threatening hemolytic uremic colitis (HUS). We examined 868 samples was taken from bovine feces and carcass from January to December 2008 in Seoul. Twenty two (9.5%) shiga toxin -producing Escherichia coli were isolated from the 230 of bovine feces, and two (0.31%) were isolated from the 638 of carcasses. Serotype of E. coli isolates were O157 (10, 41.6%), O26 (10, 41.6%), O111 (1, 4.2%) and UT (3, 12.6%). In PCR, the isolates displayed three different stx gene combination (stx1 [2, 8.4%]), stx2 [3, 12.6%] and stx1 and stx2 [19,87.5%]). The eaeA and hlyA gene were found in 11 (45.8%) of the 24 strains. Saa gene was present only one strains (4.2%). Toxin typing using reverse passive latex agglutination test showed the same result in VT 1. But it showed different result in VT 2. In antimicrobial susceptibility test, all isolates were sensitive to amikacin, amoxicillin/clavulanic acid, ciprofloxacin and colistin. Eighteen strains (75.0%) of 24 isolates showed the multi-resistant patterns with over 3 drugs. PFGE was performed after the genomic DNA of twenty four isolates was digested with Xba I. the 24 isolates showed 7 (A~G) PFGE type.

Antimicrobial Activity of Lentinus edodes Extract (표고버섯(Lentinus edodes)추출물의 항균활성)

  • 김용두;김경제;조덕봉
    • Food Science and Preservation
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    • v.10 no.1
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    • pp.89-93
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    • 2003
  • To develope natural food preservatives, ethanol and water extracts were prepared from the Lentinus edodes and antimicrobial activities were examined against 10 microorganisms which were food borne pathogens and / or food poisoning microorganisms and food-related bacteria and yeasts. Ethanol extract exhibited antimicrobial activities for the microorganisms tested, but not on lactic acid bacteria and yeast Especially, minimum inhibitory concentration(MIC) for Escherichia coli were as low as 0.5 mg/mL. Antimicrobial activity of the ethanol extract was stable by the heating at 121$^{\circ}C$ for 15 min and not affected by pH. The ethanol extract of Lentinus edodes exhibiting high antimicrobial activity. The highest antimicrobial activity adjust bacteria tested was found in the ethylacetate fraction.