• Title/Summary/Keyword: OmpW

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Regulation of an Outer Membrane Protein, OmpW, Expression and Its Biological Function in Salmonell typhimurium (Salmonella typhimurium 외막 단백질 OmpW의 발현조절 및 기능에 관한 연구)

  • Yoo, Ah-Young;Yu, Jong-Earn;Yang, Ji-Seon;Kim, Young-Hee;Bae, Chang-Ho;Oh, Jeong-Il;Kang, Ho-Young
    • Journal of Life Science
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    • v.18 no.11
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    • pp.1606-1611
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    • 2008
  • Outer membrane proteins (OMPs) expressed in the Gram negative bacteria such as Salmonella play multiple functions including material transports, adhesive factors and reception of external signals. This study has been focused on an OmpW protein known as a protein required to form a hydrophobic porin in outer membrane. We have constructed a S. typhimurium CK10 mutant deleting an ompW gene on chromosome. The CK10 strain was more tolerant to SDS than the wild-type strain did. As increase of salt concentration in the culture media, significantly decreased amount of OmpW protein in cells were detected. The maximum OmpW protein was expressed in the absence of salt supplement. However, the growth of CK10 strain was indistinguishable compared to that of the wild-type strain at the variable osmotic conditions. The biological role of differential OmpW expression in response to osmotic conditions remains to be investigated.

Genetic Variations of Outer Membrane Protein Genes of Vibrio harveyi Isolated in Korea and Immunogenicity of OmpW in Olive Flounder, Paralichthys olivaceus (한국에서 분리된 Vibrio harveyi 외막단백질의 유전적 차이와 넙치(Paralichthys olivaceus)에 대한 OmpW의 면역원성 분석)

  • KIM, Myoung-Sug;JIN, Ji-Woong;JUNG, Sung-Hee;SEO, Jung-Soo;HONG, Suhee
    • Journal of Fisheries and Marine Sciences Education
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    • v.27 no.5
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    • pp.1508-1521
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    • 2015
  • Vibrio harveyi is a pathogenic marine bacterium causing systemic symptoms resulting in mass mortalities in fishes and shrimps in aquaculture. Outer membrane proteins(OMPs) are related to the pathogenicity and thus good targets for diagnosis and vaccination for Gram negative bacteria. Recently vaccination strategies using the OMPs have been suggested to control vibriosis in several fish species. In this study, we have isolated V. harveyi from diseased marine fishes from different regions of Korea and investigated genetic variations of four OMP genes including OmpK, OmpU, OmpV and OmpW. Consequently, OmpK and U genes could be divided into 3 subgroups of type I, II, III and type A, B, C, respectively, without any correlation with geographical regions and species while OmpV and W were highly homologous. OmpW gene of V. harveyi FP4138 was fully sequenced and predicted the deduced amino acid sequence to form ${\beta}-barrel$ with hydrophobic channel. Indeed, the immunogenicity of recombinant OmpW produced in Escherichia coli was assessed by vaccinating flounder. As a result, the high antibody response with antibody titer of $4.2{\pm}0.7$ and protection with relative percent survival of 60% against artificial infection of V. harveyi were demonstrated. This result indicates that OmpW is a virulence related factor and it can be a vaccine candidate to prevent a high mortality caused by V. harveyi infection in olive flounder, Paralichthys olivaceus.

Constitutive Expression of Lipase on the Cell Surface of Escherichia coli using OmpC Anchoring Motif

  • Lee, Seung Hwan;Lee, Sang Yup
    • Korean Chemical Engineering Research
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    • v.58 no.2
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    • pp.280-285
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    • 2020
  • We have developed a constitutive display system of the Pseudomonas fluorescens SIK W1 TliA lipase on the cell surface of Escherichia coli using E. coli outer membrane protein C (OmpC) as an anchoring motif, which is an economical compared to induced system. For the constitutive expression of truncated OmpC-TliA fusion proteins, gntT104 promoter was employed. Cell growth was not affected by over expression of fusion protein during entire culture time, suggesting cell lysis was not a problem. The localization of truncated OmpC-TliA fusion protein on the cell surface was confirmed by immunofluorescence microscopy and measuring whole cell lipase activity. Constitutively displayed lipase was very stable, retaining activity enantioselectivity throughout the five repeated reactions. These results suggest that OmpC from E. coli be a useful anchoring motif for displaying enzymes on the cell surface without any inducers, and this stable surface display system can be employed for a broad range of biotechnological applications.

Proteomic Analysis of Outer Membrane Proteins in Salmonella enterica Enteritidis

  • Cho, Youngjae;Park, Soyeon;Barate, Abhijit Kashinath;Truong, Quang Lam;Han, Jang Hyuck;Jung, Cheong-Hwan;Yoon, Jang Won;Cho, Seongbeom;Hahn, Tae-Wook
    • Journal of Microbiology and Biotechnology
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    • v.25 no.2
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    • pp.288-295
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    • 2015
  • Salmonella enterica serovar Enteritidis is the predominant agent causing salmonellosis in chickens and other domestic animals. In an attempt to identify antigenic S. Enteritidis outer membrane proteins (OMPs) that may be useful for subunit vaccine development, we established a proteomic map and database of antigenic S. Enteritidis OMPs. In total, 351 and 301 spots respectively from S. Enteritidis strain 270 and strain 350 were detected by two-dimensional gel electrophoresis. Fifty-one antigen-reactive spots were detected by antisera on two-dimensional immunoblots and identified as 12 specific proteins by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. OmpA and DNA starvation/stationary phase protection protein (Dps) were the most abundant proteins among the identified OMPs, comprising 22 and 12 protein species, respectively. Interestingly, we found that the Dps of S. Enteritidis is also antigenic. OmpW was also verified to have high antigenicity. These results show that OmpA, Dps, and possibly OmpW are antigenic proteins. This study provides new insights into our understanding of the immunogenic characteristics of S. Enteritidis OMPs.

Antibacterial Effect of Various Fermentation Products and Identification of Differentially Expressed Genes of E.coli (다양한 발효액의 항균효과와 대장균의 유전적 변화에 미치는 영향)

  • Heo, Jihye
    • Korean Journal of Clinical Laboratory Science
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    • v.54 no.2
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    • pp.119-124
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    • 2022
  • Pseudomonas aeruginosa (P. aeruginosa), Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) are typical opportunistic pathogens. Moreover, these bacteria are known to possess multidrug-resistant (MDR) properties. This study investigates the antimicrobial activity of six fermented products, which have varying efficacies against P. aeruginosa, E. coli, and S. aureus. To identify novel candidate genes, differential expression analysis was performed using an annealing control primer. In the disk diffusion method, Fig vinegar (FV) and Diospyros kaki Thunb vinegar (DTV) showed the greatest increase in inhibition compared to other fermented products, whereas fermented Korean traditional nature herb (FKTNH) had no antibacterial effect. This study identified down-regulation of Escherichia coli O157:H7 ompW gene for outer membrane protein W, whereas gene for synthetic construct Lao1 gene for L-amino acid oxidase were up-regulated in E. coli treated with 5% FV. Consuming fermented vinegar helps prevent bacterial infections. Especially, FV and DTV are potentially useful alternative natural products for multidrug resistance. Furthermore, both are expected to be used as effective natural antimicrobial agents, such as disinfectants.

Millimeter-Wave(W-Band) Forward-Looking Super-Resolution Radar Imaging via Reweighted ℓ1-Minimization (재가중치 ℓ1-최소화를 통한 밀리미터파(W밴드) 전방 관측 초해상도 레이다 영상 기법)

  • Lee, Hyukjung;Chun, Joohwan;Song, Sungchan
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.28 no.8
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    • pp.636-645
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    • 2017
  • A scanning radar is exploited widely such as for ground surveillance, disaster rescue, and etc. However, the range resolution is limited by transmitted bandwidth and cross-range resolution is limited by beam width. In this paper, we propose a method for super-resolution radar imaging. If the distribution of reflectivity is sparse, the distribution is called sparse signal. That is, the problem could be formulated as compressive sensing problem. In this paper, 2D super-resolution radar image is generated via reweighted ${\ell}_1-Minimization$. In the simulation results, we compared the images obtained by the proposed method with those of the conventional Orthogonal Matching Pursuit(OMP) and Synthetic Aperture Radar(SAR).

Flame-retardant Coating of Cotton Fabric via UV curing of DMEP

  • Jang, Jin-Ho;Kim, Young-Min;Jeong, Yong-Kyun;Nam, Young Sik
    • Textile Coloration and Finishing
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    • v.19 no.5
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    • pp.1-7
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    • 2007
  • A new UV-curable coating was carried out to impart the flame-retardant property to cotton fabric using Bis[2-(methacryloyloxy)ehtyl]phosphate(DMEP) and 2-hydroxy-2-methyl-1-phenyl-propan-l-one(HMPP) as a UV-curable monomer and a photoinitiator, respectively. The cotton fabrics were applied with various amounts of DMEP and HMPP via padding and UV irradiation, and the optimal concentrations of DMEP and HMPP in the formulation were 40%(w/w) and 7%omp respectively. A limiteed oxygen index(LOI) up to 28.9 was obtained for the UV-cured fabric, which may occur through a condensed phase mechanism as verified by the increased residue number with increased application level. The characterization of the UV-cured coating was made by IR analysis, thermo-gravimetric analysis, LOI test, and scanning electron microscopy.

Versatile Catabolic Properties of Tn4371-encoded bph Pathway in Comamonas testosteroni (Formerly Pseudomonas sp.) NCIMB 10643

  • Kim, Jong-Soo;Kim, Ji-Hyun;Ryu, Eun-Kyeong;Kim, Jin-Kyoo;Kim, Chi-Kyung;Hwang, In-Gyu;Lee, Kyoung
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.302-311
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    • 2004
  • Comamonas testosteroni (formerly Pseudomonas sp.) NCIMB 10643 can grow on biphenyl and alkylbenzenes $(C_2-C_7)$ via 3-substituted catechols. Thus, to identify the genes encoding the degradation, transposon-mutagenesis was carried out using pAG408, a promoter-probe mini-transposon with a green fluorescent protein (GFP), as a reporter. A mutant, NT-1, which was unable to grow on alkylbenzenes and biphenyl, accumulated catechols and exhibited an enhanced expression of GFP upon exposure to these substrates, indicating that the gfp had been inserted in a gene encoding a broad substrate range catechol 2,3-dioxygenase. The genes (2,826 bp) flanking the gfp cloned from an SphI-digested fragment contained three complete open reading frames that were designated bphCDorfl. The deduced amino acid sequences of bphCDorfl were identical to 2,3-dihydroxybiphenyl 1,2-dioxygenase (BphC), 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase (BphD), and OrfI, respectively, that are all involved in the degradation of biphenyl/4-chlorobiphenyl (bph) by Ralstonia oxalatica A5. The deduced amino acid sequence of the orfl revealed a similarity to those of outer membrane proteins belonging to the OmpW family. The introduction of the bphCDorfl genes enabled the NT-l mutant to grow on aromatic hydrocarbons. In addition, PCR analysis indicated that the DNA sequence and gene organization of the bph operon were closely related to those in the bph operon from Tn4371 identified in strain A5. Furthermore, strain A5 was also able to grow on a similar set of alkylbenzenes as strain NCIMB 10643, demonstrating that, among the identified aromatic hydrocarbon degradation pathways, the bph degradation pathway related to Tn4371 was the most versatile in catabolizing a variety of aromatic hydrocarbons of mono- and bicyclic benzenes.

Development of Standard Analysis Methods for Physical Properties on Korean Bedsoil 2. Water content, Water retention, Saturated hydraulic conductivity (우리나라 상토의 물리적 표준분석법 설정 연구 2. 수분함량, 보수력, 포화수리전도도)

  • Kim, Lee-Yul;Jung, Kang-Ho;Ro, Hee-Myong
    • Korean Journal of Soil Science and Fertilizer
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    • v.35 no.6
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    • pp.335-343
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    • 2002
  • Methods of bedsoil analysis were difficult to be applied universally because use and material of bedsoil are diverse from country to country. Korean Standard Methods for Bedsoil Analysis was developed to measure the water content, water retention, and saturated hydraulic conductivity. Fifty-three samples for horticultural bedsoil and nine samples for paddy rice bedsoil in the current market were collected. Water content of bedsoil was determined using gravimetric method through $105^{\circ}C$ oven-dry for 16 hours, but different calculations between horticultural and paddy rice bedsoils were chosen according to different predominant component, plant residue or mineral. Water content percentage of horticultural bedsoil was calculated as [(weight of sample before oven-dry - weight of sample after oven-dry)/(weight of sample before oven-dry)]${\times}100$, while that of paddy rice bedsoil as [(weight of sample before oven-dry - weight of sample after oven-dry)/(weight of sample after oven-dry)]${\times}100$. Water retention was measured at water potential -0.5, -1, -3, -5, -7, -10 kPa by Sandbox method and saturated hydraulic conductivity was measured by constant head method using acryl cylinder (${\Phi}5cm{\times}L\;20cm$). By new 'Korean Standard Methods of Bedsoil Analysis', the average water content of horticultural bedsoil was obtained 46.34%(w/w) and that of paddy rice bedsoil 16.89%. For horticultural bedsoil, easily available water(EAW), water buffering capacity(WBC), and optimal matric potential(OMP) was estimated at 28.4%, 7.01%, and -5.60 kPa, respectively. Optimal moisture content was 44.41% and average saturated hydraulic conductivity for bedsoils was estimated at $28.4cm\;min^{-1}$.