• Title/Summary/Keyword: ompA

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Expression of Newer Outer Membrane Proteins (OMPs) Induced by Cephalosporins and Quinolone Group of Antibiotics in Klebsiella pneumoniae

  • KY TO;DANA VAN;SHARMA SAROJ;CHHIBBER SANJAY
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.421-424
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    • 2005
  • Effect of antibiotics belonging to three different groups, including third generation cephalosporins, aminoglycosides, and quinolones, on the outer membrane protein (OMP) profile of Klebsiella pneumoniae was examined. It was found that a new OMP (porins) of 40 kDa molecular mass was expressed in Klebsiella pneumoniae, when grown in the presence of ceftazidime, whereas new proteins with 30 kDa and 22 kDa masses were detected in the presence of ofloxacin. The immunoblot analysis showed that the new proteins of 40 kDa and 30 kDa molecular masses were expressed on the outer envelope, when being exposed to antibiotics ceftazidime and ofloxacin, respectively. This finding is important, as the outer surface comes in contact with the immune system, and therefore may have a bearing on the outcome of the disease.

Clinico-pathological Features of Chlamydophila psittaci Infection in Parrots and Genetic Characterization of the Isolates (Chlamydophila psittaci에 감염된 앵무새의 임상병리학적 특징과 원인균의 ompA 유전자 비교분석)

  • Kim, Il-Hwan;Jang, Jin-Wook;Lee, Su-Hyung;Kim, Dae-Yong;Seong, Won-Jin;Kwon, Hyuk-Joon;Kim, Jae-Hong
    • Korean Journal of Veterinary Research
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    • v.52 no.3
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    • pp.193-198
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    • 2012
  • Avian chlamydiosis is caused by Chlamydophila psittaci and considered as one of an important zoonotic disease throughout the world. Among more than 400 avian species including poultry and pet birds susceptible to the disease, psittacine birds were known to be mostly susceptible hosts. In Korea, no outbreak of the disease and genetic analysis of the agent in poultry and pet birds have been reported. With histopathological findings and genetic identification of a causative agent, avian chlamydiosis was identified in parrots submitted from the same pet bird farm in 2006 and 2009 for the diagnosis. Based on genetic sequences and phylogenetic analysis of ompA gene, the two isolates of Chlamydophila psittaci showed 100% of genetic similarity and belonged to genotype A, suggesting that the same agent might be continuously circulated in the farm. This result indicates that serological survey of the disease in pet bird farms and impact of the disease on significance in public health may be further studied.

Proteomic Analysis and Protective Effects of Outer Membrane Proteins from Salmonella Gallinarum in Chickens (Salmonella Gallinarum 세포외막단백질의 프로테옴 분석 및 닭에서의 방어능 효과)

  • Sun, Jisun;Cho, Youngjae;Jang, Joo-Hyun;Kang, Zheng-Wu;Han, Jang-Hyuk;Hahn, Tae-Wook
    • Food Science of Animal Resources
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    • v.33 no.2
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    • pp.281-286
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    • 2013
  • Salmonella Gallinarum (SG) is known as an important pathogen that causes fowl typhoid in chickens. To investigate SG outer-membrane proteins (OMPs) as a vaccine candidate, we used proteomic mapping and database analysis techniques with extracted OMPs. Also, extracted OMPs were evaluated in several aspects to their safety, immune response in their host and protective effects. Our research has established a proteomic map and database of immunogenic SG-OMPs used as inactive vaccine against salmonellosis in chickens. A total of 22 spots were detected by 2-dimensional gel electrophoresis and immunogenic protein analysis. Eight spots were identified by Matrix-Assisted Laser Desorption/Ionization-Time of Flight-Mass spectrometry (MALDI-TOF-MS) and peptide mass fingerprinting (PMF) and categorized into four different types of proteins. Among these proteins, OmpA is considered to be an immunogenic protein and involved in the hosts' immune system. To estimate the minimum safety dose in chickens, 35 brown layers were immunized with various concentrations of OMPs, respectively. Consequently, all chickens immunized with more than a $50{\mu}g$ dose were protected against challenges. Moreover, intramuscular administration of OMPs to chickens was more effective compared to subcutaneous administration. These results suggest that the adjuvanted SG-OMP vaccine not only induces both the humoral and cellular immune response in the host but also highly protects the hosts' exposed to virulent SG with $50{\mu}g$ OMPs extracted by our method.

Multi-Immunogenic Outer Membrane Vesicles Derived from a MsbB-Deficient Salmonella enterica Serovar Typhimurium Mutant

  • Lee, Sang-Rae;Kim, Sang-Hyun;Jeong, Kang-Jin;Kim, Keun-Su;Kim, Young-Hyun;Kim, Sung-Jin;Kim, E-Kyune;Kim, Jung-Woo;Chang, Kyu-Tae
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1271-1279
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    • 2009
  • To develop low endotoxic and multi-immunogenic outer membrane vesicles (OMVs), a deletion mutant of the msbB gene in Salmonella enterica serovar Typhimurium (S. Typhimurium) was used as a source of low endotoxic OMV, and an expression vector of the canine parvovirus (CPV) VP2 epitope fused to the bacterial OmpA protein was constructed and transformed into the Salmonella ${\Delta}msbB$ mutant. In a lethality test, BALB/c mice injected intraperitoneally with the Salmonella ${\Delta}msbB$ mutant survived for 7 days, whereas mice injected intraperitoneally with the wild type survived for 3 days. Moreover, all mice inoculated orally with the ${\Delta}msbB$ mutant survived for 30 days, but 80% of mice inoculated orally with the wild type survived. The OmpA::CPV VP2 epitope fusion protein was expressed successfully and associated with the outer membrane and OMV fractions from the mutant S. Typhimurium transformed with the fusion protein-expressing vector. In immunogenicity tests, sera obtained from the mice immunized with either the Salmonella msbB mutant or its OMVs containing the OmpA::CPV VP2 epitope showed bactericidal activities against wild-type S. Typhimurium and contained specific antibodies to the CPV VP2 epitope. In the hemagglutination inhibition (HI) assay as a measurement of CPV-neutralizing activity in the immune sera, there was an 8-fold increase of HI titer in the OMV-immunized group compared with the control. These results suggested that the CPV-neutralizing antibody response was raised by immunization with OMV containing the OmpA::CPV VP2 epitope, as well as the protective immune response against S. Typhimurium in BALB/c mice.

Comparative Phenotypic Analysis of Anabaena sp. PCC 7120 Mutants of Porin-like Genes

  • Schatzle, Hannah;Brouwer, Eva-Maria;Liebhart, Elisa;Stevanovic, Mara;Schleiff, Enrico
    • Journal of Microbiology and Biotechnology
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    • v.31 no.5
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    • pp.645-658
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    • 2021
  • Porins are essential for the viability of Gram-negative bacteria. They ensure the uptake of nutrients, can be involved in the maintenance of outer membrane integrity and define the antibiotic or drug resistance of organisms. The function and structure of porins in proteobacteria is well described, while their function in photoautotrophic cyanobacteria has not been systematically explored. We compared the domain architecture of nine putative porins in the filamentous cyanobacterium Anabaena sp. PCC 7120 and analyzed the seven candidates with predicted OprB-domain. Single recombinant mutants of the seven genes were created and their growth capacity under different conditions was analyzed. Most of the putative porins seem to be involved in the transport of salt and copper, as respective mutants were resistant to elevated concentrations of these substances. In turn, only the mutant of alr2231 was less sensitive to elevated zinc concentrations, while mutants of alr0834, alr4741 and all4499 were resistant to high manganese concentrations. Notably the mutant of alr4550 shows a high sensitivity against harmful compounds, which is indicative for a function related to the maintenance of outer membrane integrity. Moreover, the mutant of all5191 exhibited a phenotype which suggests either a higher nitrate demand or an inefficient nitrogen fixation. The dependency of porin membrane insertion on Omp85 proteins was tested exemplarily for Alr4550, and an enhanced aggregation of Alr4550 was observed in two omp85 mutants. The comparative analysis of porin mutants suggests that the proteins in parts perform distinct functions related to envelope integrity and solute uptake.

Comparison of Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, and Loop-Mediated Isothermal Amplification for the Detection of Cronobacter sakazakii in Milk Powder (분유에 오염된 Cronobacter sakazakii 검출을 위한 중합효소연쇄반응, 실시간중합효소연쇄반응, 등온검출법의 비교)

  • Kim, Young-Joo;Seo, Sheungwoo;Wang, Xiaoyu;Seo, Dong Joo;Lee, Min Hwa;Son, Na Ry;Lee, Bog-Hieu;Choi, Changsun
    • Food Science of Animal Resources
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    • v.33 no.5
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    • pp.610-616
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    • 2013
  • Loop-mediated isothermal amplification (LAMP) is an emerging detection technology for the amplification of DNA under isothermal conditions. The aim of this study was to develop a rapid and reliable LAMP technique for the detection of Cronobacter sakazakii in milk powder. In order to enhance the sensitivity and specificity, LAMP primers targeting outer membrane protein A (ompA) gene of C. sakazakii were designed using Explorer V4 software. Thirty seven C. sakazakii strains and 13 pathogenic microorganisms were used for comparative detection of C. sakazakii using polymerase chain reaction (PCR), real-time PCR, and LAMP. LAMP developed in this study could specifically detect C. sakazakii strains without cross-reactivity with other foodborne pathogens. LAMP products amplified from ompA gene of C. sakazakii were digested with with HhaI and NruI enzyme. The specificity of LAMP was confirmed by restriction fragment length polymorphism (RFLP) analysis. LAMP could detect C. sakazakii within 1 h without bacterial culture and its detection limit was as low as 1 CFU/mL C. sakazakii in milk. In the comparison of the sensitivity, LAMP showed 10,000- and 100-times higher detection limit than PCR or real-time PCR, respectively. Therefore, this study can conclude that LAMP is a rapid and reliable detection technique for C. sakazakii contaminated in powdered milk.

Study of the Compressed and Expanded Liquid Rubidium with the Optimized Nonlocal Model Potential

  • 강홍석;박성호
    • Bulletin of the Korean Chemical Society
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    • v.20 no.11
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    • pp.1288-1294
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    • 1999
  • We show that Shaw's optimized nonlocal model potential (OMP) in combination with the perturbative hyper-netted-chain equation for pair correlation functions can be successfully applied to predict pair structures of compressed and expanded liquid rubidium. For compressed rubidium, it is possible to apply the OMP to a state for which the model radius is even close to the Wigner-Seitz radius. In addition, our results are parallel to those from Chihara and Kahl's quantal hypernetted-chain equation in that it supports the uniform compression model up to 6.1 GPa. Calculation also shows that the pair structure is relatively insensitive to the choice of the exchange-correlation function for the electron liquid. Discussions are also given for compressed and expanded cesium.

Physicochemical Properties and the Origin of Summer Bottom Cold Waters in the Korea Strait (하계 대한해협 저층냉수의 물리.화학적인 특성 및 기원)

  • Kim, Il-Nam;Lee, Tong-Sup
    • Ocean and Polar Research
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    • v.26 no.4
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    • pp.595-606
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    • 2004
  • Hydrographic survey in the Korea Strait has long history that has begun in August 1917 at the Busan - Tsushima cross section, still continues to date. However, chemical properties of bottom cold water found exclusively in the western channel of the Korea Strait during summer did not receive much scientific attention. The aim of the study is to decipher the enigmatic origin of the Korea Strait Bottom Cold Water (KSBCW) in terms of chemical properties. The physicochemical properties of the KSBCW are extracted from the CREAHS II hydrographic data. OMP method was applied to analyze origin of the KSBCW quantitatively. The KSBCW is well defined by low temperature below $10^{\circ}C$. The cold waters exhibited the local presence near the coast at about 120m depth with a thickness of 20m to 30m. The cold water was characterized by relatively cold, saline and higher chemical concentrations than adjacent waters. The KSBCW seems to have different origin kom that of the coastal upwelled waters at the Ulgi-Gampo because it is saline, denser and contains considerably less dissolved oxygen than upwelled waters. The physicochemical properties are reported to have noticeable annual variations which suggest the complex origin of the KSBCW. OMP analysis show that the KSBCW is a mixture of three water types; TMW (24%), ESIW (36%) and ESPW (40%). Relationship between the KSBCW and the east Sea circulation is traced by mapping the water masses that have similar T, S and DO of KSBCW. The result showed that the KSBCW is most possibly an extension of southward flowing coastal intermediate waters. Front these results, we expect that the monitoring KSBCW will provide us valuable information about the East Sea circulation.

Comparison of diagnostic methods for detection of Brucella species in dog blood samples (개 혈액 재료에서의 Brucella 검출을 위한 진단방법의 비교)

  • Kwon, Soon-Oh;Lam, Truong Quang;Her, Moon;Ahn, Dong-Chun;Park, Sang-Hee;Park, Mi-Yeoun;Lee, Young-Ju;Hahn, Tae-Wook
    • Korean Journal of Veterinary Service
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    • v.32 no.4
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    • pp.335-341
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    • 2009
  • Canine brucellosis produce abortions and infertility in dogs and is currently diagnosed by serological methods such as rapid slide agglutination test with 2-mercaptoethanol (2-ME RSAT) and immunochromatographic assay (ICA). Bacterial isolation is considered gold standard for Brucella diagnosis and the polymerase chain reaction (PCR) is an alternative method to bacterial isolation. A total of 36 whole blood samples were collected from dogs reared in area of Chuncheon and were subjected to serology (2-ME RSAT and ICA for B. canis, Rose Bengal test and C-ELISA for B. abortus), blood culture and 3 types of PCRs (BSCP31, 16s rRNA, and OMP-2). All blood samples were negative by serology and blood cultures. The BCSP31 and the OMP-2 PCR detected 5 samples were positive whereas the 16S rRNA PCR detected all samples were negative as serological methods and blood culture did. From the results observed in the present study, we conclude that 16S rRNA PCR could be used for direct PCR for canine blood samples.

Heavy Metal Detection and Removal in Artificial Wastewater Using Two-Component System Based Recombinant Bacteria (Two-component System 기반 재조합균을 이용한 인공폐수에서의 중금속 인지 및 제거)

  • Ravikumar, Sambandam;Hong, Soon-Ho;Yoo, Ik-Keun
    • Korean Journal of Microbiology
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    • v.48 no.3
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    • pp.187-191
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    • 2012
  • Two-component system (TCS)-based bacterial zinc and copper biosensors, in which green fluorescent protein (GFP) is expressed under the control of zraP and cusC promoter in ZraS/R and CusS/R TCS, were evaluated in artificial wastewater. Bacterial biosensors developed in this study efficiently expressed GFP by the recognition of $Zn^{2+}$ and $Cu^{2+}$ in artificial wastewater. Secondly, TCS-based zinc and copper removing bacteria with the peptide displayed on cell surface were examined in artificial wastewater. Zinc and copper removing bacteria expressed the peptide as a fusion protein such as OmpC-ZBP (zinc binding peptide) and OmpC-CBP (copper binding peptide) on the cell surface when sensing exogenous $Zn^{2+}$ and $Cu^{2+}$ through ZraS/R and CusS/R TCS. The recombinant cell expressing metal-adsorbing peptide could efficiently remove copper and zinc (15 and 18 mg/g dry cell weight, respectively) in artificial wastewater. Therefore, it was demonstrated that the TCS-based recombinant cell for the recognition or removal of heavy metal functions well in artificial wastewater environment.