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Temperature-Dependency Urease Activity in Vibrio parahaemolyticus is Related to Transcriptional Activator UreR

  • Park, Kwon-Sam;Lee, Soo-Jae;Chung, Yong-Hyun;Iida, Tetsuya;Honda, Takeshi
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1456-1463
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    • 2009
  • Vibrio parahaemolyticus possessing urease-positive property is relatively rare, but such strains consistently exhibit the TDH-related hemolysin (TRH) gene. In this study, we examined the effects of incubation temperature on urease activity expression, using the TH3996 and AQ4673 strains where the enzyme activity is known to be temperature-dependent and -independent, respectively. In the TH3996 strain, $\beta$-galactosidase activity was 4.4-fold lower after $30^{\circ}C$ cultivation than after $37^{\circ}C$ in a ureR-lacZ fusion strain, but temperature dependency was not found in ureD- or nikA-lacZ fusion strains. However, ureR-, ureD-, and nikA-lacZ fusions of the AQ4673 strain was not influenced by incubation temperature. We compared the promoter sequences of ureR between the above two strains. Intriguingly, we detected mismatches of two nucleotides between the two strains located at positions -66 and -108 upstream of the methionine initiation codon for UreR. Additionally, urease activity was not affected by culture temperature at either $30^{\circ}C$ or $37^{\circ}C$ by allelic introduction of the AQ4673 ureR gene into the TH3996 ureR deletion mutant. Taken together, our study demonstrates that the transcriptional factor UreR is involved in the temperature dependency of urease activity, and two nucleotides within the ureR promoter region are of particular importance for the urease activity dependency of V. parahaemolyticus.

Purification, Characterization and Cellular Localization of Klebsiella aerogenes UreG Protein

  • Lee, Mann-Hyung
    • Biomolecules & Therapeutics
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    • v.3 no.4
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    • pp.311-315
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    • 1995
  • The K. aerogenes ureal gene product was previously shown to facilitate assembly of the crease metallocenter (Lee, M. H., Mulrooney, S. B., Renner, M. J., Markowicz, Y., and Hausinger, R. P. (1992) J. Bacteriol. 174, 4324-4330). UreG protein has now been purified and characterized. Although the protein is predicted to possess a putative NTP-binding P-loop motif, equilibrium dialysis studies showed negative results. Immunogold electron microscopic studies using polyclonal antibodies directed against UreG protein confirm that UreG is located in the cytoplasm as predicted in the DNA sequence.

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Separate Expression and in vitro Activation of Recombinant Helicobacter pylori Urease Structural Subunits

  • Lee, Kwang-Kook;Son, Joo-Sun;Chang, Yung-Jin;Kim, Soo-Un;Kim, Kyung-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.8 no.6
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    • pp.700-704
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    • 1998
  • Each of the recombinant structural genes of Helicobacter pylori urease, ureA and ureB, was cloned and overexpressed as inclusion bodies. Solubilization and renaturation of the inclusion bodies were carried out, to accelerate the pairing of sulfhydryl groups and the incorporation of nickel ions, which would lead to the native structure with high enzyme activity. Rates of urea hydrolysis were monitored as an indication of in vitro activation of renatured ureases. The activation of the apoprotein using 1 mM nickel ion, 100 mM sodium bicarbonate and a 10:1 ratio of reducing power resulted in a weak urease activity (about 11% of the native urease activity encoded by pTZ 19R/ure-l). When a sparse matrix screen method originally discovered for the crystallization of proteins was used, the activity increased higher than that obtained using glutathione. The effect of polyethylene glycol (PEG) on the activity was noticeable, giving two-fold increase in the specific activity (about 11 U/mg of protein corresponding to 22% of the native urease activity encoded by pTZ19R/ure-1).

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Isolation and Identification of Urease-Positive Photobacterium sp. Strain HA-2 from Sea Water (해수에서 urease 양성 Photobacterium sp. Strain HA-2의 분리 및 동정)

  • Kim, Kang-Jin;No, A-Reum;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.42 no.6
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    • pp.531-536
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    • 2009
  • A urease-positive bacterium isolated from sea water was identified as Photobacterium sp. by morphological, biochemical, and 16s rRNA gene analyses and named Photobacterium sp. strain HA-2. 2.0-fold increase enzyme activity was observed in LB medium containing 3% NaCl and 0.1% urea or not and the enzyme activity was 16.0-fold lower compared to urease-positive Vibrio parahaemolyticus AQ4673 strain when grown in the LB medium containing 3% NaCl with 0.1% urea. The cloning and sequencing of Photobacterium sp. strain HA-2 urease gene cluster is currently being analyzed in our laboratory.

Electrical Characteristics of ($Sr_{0.85}Ca_{0.15})TiO_3$ Thin Film by RF Sputtering Method (RF 스퍼터링법에 의한 ($Sr_{0.85}Ca_{0.15})TiO_3$ 박막의 전기적 특성)

  • 장원석;김진사;최운식;김충혁;이준웅
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 1998.11a
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    • pp.239-242
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    • 1998
  • The (S $r_{0.85}$C $a_{0.15}$)Ti $O_3$ (SCT) thin films are deposited on Pt-coated electrode using RF magnetron sputtering method at various substrate temperature. Dielectric constant of SCT thin films is increased with increased as the deposition temperature and changes almost linearly in temperature ranges from -80 to +90[$^{\circ}C$]. The drastic decrease of dielectric constant and increase of dielectric loss in SCT thin films is observed above 200[kHz]. V-I characteristics of SCT thin films show the increasing leakage current with the increases of deposition temperature.ure.

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The Structural and Electrical Properties of Vanadium Oxide Thin Films as $O_2/(Ar+O_{2})$ ratio ($O_2/(Ar+O_{2})$비에 따른 바나듐 산화막의 구조적, 전기적 특성)

  • 최용남;최복길;최창규;김성진
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2001.07a
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    • pp.729-732
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    • 2001
  • In this study, the effect of oxygen partial pressure on the electrical properties of vanadium oxide(VO$_{x}$) thin films were investigated. The thin films were prepared by r.f. magnetron sputtering from V$_2$O$_{5}$ target in a gas mixture of argon and oxygen. The oxygen partial pressure ratio is changed from 0% to 8%. I-V characteristics were distinguished between linear and nonlinear region. In the low field region the conduction is due to Schottky emission, while at high fields it changes to Fowler-Nordheim tunneling type conduction. The conductivity measurements have shown an Arrhenius dependence of the conductivity on the temperature.ure.

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Isolation of Photobacterium damselae subsp. damselae from zebra shark Stegostoma fasciatum

  • Han, Jee Eun;Gomez, Dennis K.;Kim, Ji Hyung;Choresca, Casiano H. Jr.;Shin, Sang Phil;Baeck, Gun Wook;Park, Se Chang
    • Korean Journal of Veterinary Research
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    • v.49 no.1
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    • pp.35-38
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    • 2009
  • The zebra shark Stegostoma fasciatum which had been reared in the commercial aquaria was found dead and submitted for postmortem examination. A pure bacterial culture was isolated from pale and enlarged liver. The analysis of ureC and 16S rRNA genes confirmed the isolate as Photobacterium (P.) damselae subsp. damselae and this pathogen was sensitive to gentamicin. Although, no mortality in mouse was observed in the experimental infection study, the isolation of this pathogen in aquarium fish is significant because it can act as a reservoir to other aquatic animals and can also be zoonotic potential to human during aquarium management. This paper describes the first isolation of P. damselae subsp. damselae from zebra shark.

Evaluation of Factors that Can Affect Protective Immune Responses Following Oral Immunization of Recombinant Helicobacter pylori Urease Apoenzyme

  • Kim, Jang-Seoung;Chang, Ji-Hoon;Park, Eun-Jeong;Chung, Soo-Il;Yum, Jung-Sun
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.865-872
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    • 2000
  • Helocobacter phylori is the major cause of gastritis, peptic ulcer, and a principal risk factor for gastric cancer. As the firs step towards a vaccine against H. pylori infection, Hy.pylori urease was expressed and purified as a recombinant apoenzyme (rUrease) in E. coli. In order to develop an effective immunization protocol using rUrease, the host immune responses were evaluated after the oral immunization of mice with rUrease preparations plus cholera toxin relative to various conditions, such as the physical nature of the antigen, the frequency of the booster immunization, the dose of the antigen, and the route of administration. The protective efficacy was assessed using a quantitative culture following an H. pylori SS1 challenge. It was demonstrated that rUrease, due to its particulated nature, was more superior than the UreB subunit as a vaccine antigen. The oral immunization of rUrease elicited significant systemic and secretory antibody responses, and activated predominantly Th2-type cellular responses. The bacterial colonization was significantly reduced (~100-fold) in those mice immunized with three or four weekly oran doses of rUrease plus cholera toxin (p<0.05), when compared to the non-immunized/challenged controls. The protection correlated well with the elicited secretory IgA level against rUrease, and these secretory antibody responses were highly dependent on the frequency of the booster immunization, yet unaffected by the dose of the antigen (25-200$\mu\textrm{g}$). These results demonstrate the remarkable potential of rUrease as a vaccine antigen, thereby strengthening the possibility of developing an H. pylori vaccine for humans.

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The Effect of Neurofeedback Traing on the hearing impairments Student about Emotional (뉴로피드백 훈련이 청각장애 학생들의 정서적 성향에 미치는 영향)

  • Bak, Ki-Ja;Ahn, Sang-Kyun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.12 no.9
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    • pp.3897-3903
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    • 2011
  • This study was to examine the effectiveness of neurofeedback training by observing the pre and post brainwave measurement results of about 39(experimental group 25, comparative group 14) student who have shown emotional. The study was examined at S city M dong A hearing impairments student, from Mar. 2009 and to Feb. 2010. The training took place two times a week, for about 40 minutes per session. The tool used to meas ure the psychological reaction was POMS (Profile of Mood State). First, the result confirmed the differences of both attention quotient(L R) and resistance stress quotient(L R). Second,, the results of the analysis show mood state. Therefore, the result of the study shows that there is possibility that NeuroFeedback technique might be positively affecting emotional of the hearing impairments student.

Effect of the Oxygen Partial Pressure on the Phase Stability of ${Bi_2}{Sr_2}{CaCu_2}{O_{8+x}}$ and ${Bi_2}{Sr_2}{Ca_2}{Cu_3}{O_{10+x}}$ (${Bi_2}{Sr_2}{CaCu_2}{O_{8+x}}$${Bi_2}{Sr_2}{Ca_2}{Cu_3}{O_{10+x}}$의 상 안정성에 대한 산소분압의 영향)

  • Park, Min-Su;Lee, Hwa-Seong;An, Byeong-Tae
    • Korean Journal of Materials Research
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    • v.5 no.5
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    • pp.583-597
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    • 1995
  • We investigated the effect of the oxygne partial pressure on the phase stability of B $i_{2}$S $r_{2}$Ca C $u_{2}$ $O_{8+x}$ and B $i_{2}$S $r_{2}$C $a_{2}$C $u_{3}$ $O_{10+x}$ at 82$0^{\circ}C$. As the oxygen pressure decreased, B $i_{2}$Sr/sb 2/CaC $u_{2}$ $O_{8+x}$ melted at 2.2$\times$10$^{-3}$ atm $O_{2}$. In the case of the B $i_{1.7}$P $b_{0.4}$S $r_{2}$C $a_{2}$ $O_{10+x}$, it started to decompose into theree phases of B $i_{2}$S $r_{2}$Cu $O_{6+y}$, $Ca_{2}$Cu $O_{3}$ and C $u_{4}$ $O_{3}$ and C $u_{4}$ $O_{3}$ at 8.0$\times$10$^{-3}$ atm $O_{2}$ and was completely decomposed at 4.3$\times$10$^{-3}$ atm $O_{2}$ B $i_{2}$S $r_{2}$C $a_{2}$C $u_{3}$ $O_{10+x}$ phase was not formed by the solid state reaction from the mixutre of $i_{2}$S $r_{2}$CaCu.sub 2/ $O_{8+x}$, $Ca_{2}$Cu $O_{3}$ and CuO down to 2.2$\times$10.sub -3/ atm O.sub 2/ but formed by the solidifciation of the formed from the mixture of the intermediate compounds in the Bi-Sr-Ca-Cu-O system and the fromation temperature of Bi.sub 2/S $r_{2}$C $a_{2}$Cu.$_{3}$ $O_{10+x}$ can be lowered by reducing oxygen partial pressure.e.e.ure.e.e.

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