• Title/Summary/Keyword: ompR

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Characteristics of Vibrio anguillarum Isolated from Seawater Cultured Rainbow Trout Oncorhynchus mykiss in Korea (해수 사육 무지개송어(Oncorhynchus mykiss)에서 분리된 Vibrio anguillarum의 특성 분석)

  • Chun, Hye-Jin;Kim, Wi-Sik;Cho, Mi-Young;Jung, Sung-Hee;Han, Hyun-Ja
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.3
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    • pp.254-261
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    • 2018
  • From 2014 to 2017, mortalities of seawater-cultured rainbow trout Oncorhynchus mykiss, were observed in the Goheung and Jeju areas of Korea, with Vibrio anguillarum (seven strains: RT1, 2, 3, 4, 5, 6, and 7) identified as the etiological agent. The phenotypic (based on API 20NE, API ZYM, and E-test kits), serotypic (slide agglutination tests with O1, O2, O3, O4, and O7 antisera), and genotypic (16S rRNA and ompU sequencing) characteristics of the seven RT strains were analyzed and compared to those of seven additional V. anguillarum stains (SF, isolated from sweet fish; FM, isolated from flathead mullet; ATCC43305; ATCC43311; ATCC43307; ATCC43308; and KCTC2711). The phenotypes of the RT strains showed variance, while the slide agglutination tests of the RT1-7, SF, and FM strains all showed positive reactions with serotype O1 antiserum. The 16S rRNA and ompU sequences of the RT1-7, SF, and FM strains were affiliated with V. anguillarum ATCC43305 (Serotype O1), but the ompU sequence of the SF strain differed from those of the RT1-7, FM, and ATCC43311 strains, including one amino acid substitution. We thus confirmed that serotype O1 V. anguillarum, with multiple phenotypes, continues to infect seawater-cultured rainbow trout in Korea.

Expression of the EPO-like Domains of Human Thrombopoietin in Escherichia coli

  • Koh, Yeo-Wook;Koo, Tai-Young;Ju, Sang-Myoung;Kwon, Chang-Hyuk;Chung, Joo-Young;Park, Myung-Hwan;Yang, Jai-Myung;Park, Seung-Kook
    • Journal of Microbiology and Biotechnology
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    • v.8 no.6
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    • pp.553-559
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    • 1998
  • cDNA of human thrombopoietin (hTPO) amplified by polymerase chain reaction from a cDNA library of human fetal liver was cloned. EPO-like domains ($hTPO_{153} \;or\; hTPO_{l63})\; of\; hTPO(hTPO_{332}$) were expressed in Escherichin coli using several kinds of expression systems, such as ompA secretion, thioredoxin fusion, and the $P_L$ and T7 expression systems. To obtain $hTPO_{153}$ in soluble form, $hTPO_{153}$ cDNA was fused in-frame behind the gene encoding ompA signal sequence and thioredoxin protein. When fused with either of the genes, $hTPO_{153}$ was not expressed to the detectable level. However, a high level expression of the EPO-like domain of hTPO was obtained using the PL and T7 expression system. $hTPO_{153} \;or\; hTPO_{l63} cDNA were subcloned into the pLex and pET-28a(+) vectors under the control of the inducible$ P_L\;T_7$ promoter, respectively. Proteins expressed using pl.ex vector and pET-28a(+) detected in insoluble forms with an expression level of about 14% and 9% of total cellular proteins, respectively, and the level of expression was rapidly diminished in 2 h after the maximum level of expression was reached.

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Soluble Expression of OmpA from Haemophilus parasuis in Escherichia coli and Its Protective Effects in the Mouse Model of Infection

  • Ahn, Jungoh;Hong, Minhee;Yoo, Sungsik;Lee, Eungyo;Won, Hokeun;Yoon, Injoong;Jung, Joon-Ki;Lee, Hongweon
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1307-1309
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    • 2012
  • Haemophilus parasuis causes contagious porcine Gl$\ddot{a}$sser's disease leading to severe losses in the swine industry. In this study, we established an efficient Escherichia coli-based system for the expression of H. parasuis major outer-membrane protein (MOMP) that has been known as a good vaccine candidate against Gl$\ddot{a}$sser's disease. Use of an E. coli-derived pelB leader sequence made it possible to produce recombinant MOMP (rMOMP) as the soluble forms without an additional refolding process. Using two different animal models, it was evaluated that the rMOMP was capable of inducing a significant immune response and providing protection against H. parasuis infection.

Increased Immunogenicity and Protective Efficacy of a P. aeruginosa Vaccine in Mice Using an Alum and De-O-Acylated Lipooligosaccharide Adjuvant System

  • Ryu, Ji In;Wui, Seo Ri;Ko, Ara;Do, Hien Thi Thu;Lee, Yeon Jeong;Kim, Hark Jun;Rhee, Inmoo;Park, Shin Ae;Kim, Kwang Sung;Cho, Yang Je;Lee, Na Gyong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.8
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    • pp.1539-1548
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    • 2017
  • Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen that commonly causes fatal infections in cystic fibrosis and burn patients as well as in patients who are hospitalized or have impaired immune systems. P. aeruginosa infections are difficult to treat owing to the high resistance of the pathogen to conventional antibiotics. Despite several efforts, no effective prophylactic vaccines against P. aeruginosa are currently available. In this study, we investigated the activity of the CIA06 adjuvant system, which is composed of alum and de-O-acylated lipooligosaccharide, on a P. aeruginosa outer membrane protein (OMP) antigen vaccine in mice. The results indicated that CIA06 significantly increased the antigen-specific IgG titers and opsonophagocytic activity of immune sera against P. aeruginosa. In addition, the antibodies induced by the CIA06-adjuvanted vaccine exhibited higher cross-reactivity with heterologous P. aeruginosa strains. Finally, mice immunized with the CIA06-adjuvanted vaccine were effectively protected from lethal P. aeruginosa challenge. Based on these data, we suggest that the CIA06 adjuvant system might be used to promote the immunogenicity and protective efficacy of the P. aeruginosa OMP vaccine.

Identification of Secretion Signals of Target Proteins in Salmonella enterica serovar Typhimurium and Construction of Secretion Vector using this Signal (Salmonella enterica serovar Typhimurium에서 Type III 분비장치의 표적단백질들의 분비신호 확인 및 Type III 분비장치를 이용한 Secretion Vector의 개발)

  • Choi, Hyuk-Jin;Eom, Joon-Ho;Cho, Jung-Ah;Lee, Sun;Lee, Kyoung-Mi;Lee, In-Soo;Park, Yong-Keun
    • Korean Journal of Microbiology
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    • v.36 no.4
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    • pp.254-258
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    • 2000
  • Invasion process of bacterial cell into intestinal epithelium is important in Salmonella infection. The invasion is induced by the proteins secreted by type III secretion appratus of Salmonella. It has been known that the proteins do not have N-terminal signal peptide existing in general secreted proteins. Recent studies on Yersinia reported that secretion signal of type III appratus may lie on 5'end secondary structure of mRNA of secreted protein. In this study, we constructed translational fusion of ompR and sopE, encoding type III secretion protein of Salmonella, and observed secretion of the fusion protein for investigating the secretion signal of Salmonella type III appratus. The sopE DNA fragments of the translational fusion contain the region of promoter and from start code to tenth or to fifth code. These translational fusions indicate that type III secretion signal of Salmonella is located on 5'end of mRNA encoding secreted protein. We constructed prototype of secretion vector using this signal to produce useful foreign protein.

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Intranasal and intraperitoneal immunization against Brucella infection using niosome and mannosylated niosomes containing Brucella recombinant trigger factor/Bp26/Omp31 chimeric protein in a mouse model

  • Fahimeh Sharif;Razieh Nazari;Mahdi Fasihi-Ramandi;Ramezan Ali Taheri;Mohsen Zargar
    • Clinical and Experimental Vaccine Research
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    • v.13 no.3
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    • pp.232-241
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    • 2024
  • Purpose: Brucellosis, a zoonotic infectious disease, is a worldwide health issue affecting animals and humans. No effective human vaccine and the complications caused by the use of animal vaccines are among the factors that have prevented the eradication of the disease worldwide. However, bio-engineering technologies have paved the way for designing new targeted and highly efficacious vaccines. In this regard, the study aimed to evaluate immunity induced by mannosylated niosome containing Brucella recombinant trigger factor/Bp26/Omp31 (rTBO) chimeric protein in a mouse model. Materials and Methods: rTBO as chimeric antigen (Ag) was expressed in Escherichia coli BL21 (DE3) and, after purification, loaded on niosome and mannosylated niosome. The characteristics of the nanoparticles were assessed. The mice were immunized using rTBO, niosome, and mannosylated niosome-rTBO in intranasal and intraperitoneal routes. Serum antibodies (immunoglobulin [Ig]A, IgG, IgG1, and IgG2a) and splenocyte cytokines (interferon-gamma, interleukin [IL]-4, and IL-12) were evaluated in immunized mice. Finally, immunized mice were challenged by B. melitensis and B. abortus. A high antibody level was produced by niosomal antigen (Nio-Ag) and mannosylated noisomal antigen (Nio-Man-Ag) compared to the control after 10, 24, and 38 days of immunization. The IgG2a/IgG1 titer ratio for Nio-Man-Ag was 1.2 and 1.1 in intraperitoneal and intranasal methods and lower than one in free Ag and Nio-Ag. Cytokine production was significantly higher in the immunized animal with Ag-loaded nanoparticles than in the negative control group (p<0.05). Moreover, cytokine and antibody levels were significantly higher in the injection than in the inhalation method (p<0.05). Results: The combination of mannosylated noisome and rTBO chimeric proteins stimulate the cellular and humoral immune response and produce cytokines, playing a role in developing the protective acquired immune response in the Brucella infectious model. Also, the intraperitoneal route resulted in a successful enhancement of cytokines production more than intranasal administration. Conclusion: Designing an effective vaccine candidate against Brucella that selectively induces cellular and humoral immune response can be done by selecting a suitable nanoniosome formulation as an immunoadjuvant and recombinant protein as an immune response-stimulating Ag.

Regulation of stf Operon Expression and Its Virulence (살모넬라가 발현하는 stf 오페론의 조절과 병원성 인자로서의 기능)

  • Kim Sam-Woong;Kim Young-Hee;Kang Ho-Young
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.553-560
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    • 2005
  • The stf (Salmonella typhimurium fimbriae) operon consisting of stfA(CDEFG assumes to encode putative fimbriae. The complete stf operon is existed in S. typhimurium and S. choleraesuis, whereas it is absent in S. typhi. Analyses of the amino acid residues between major subunit StfA of the Stf fimbriae and those of known other fimbriaes suggested that Stf belongs to class I type fimbriae. Through comparison of StfD chaperone with the other fimbrial chaperones, and of C-terminus in subunits of Stf fimbriae, it belongs to FGS (with a short Fl-G1 loop) subfamily. In order to investigate the expression of stf operon, we have constructed a Salmonella strain containing a chromosomal stfA::lacZYA transcriptional fusion, resulting in S. typhimurium $_X8532$. The strain $_X8532$ lacked the expression of \beta-galactosidase$ under normal culture conditions. However, with longer incubation time of the S. typhimurium $_X8532$, we have isolated 21 individual strains exhibiting $Lac^+$ phenotype. $Lac^+$ phenotype was appeared as approximately 0.03 frequency per generation. All isolates expressed lacZ constitutively in the various environmental conditions. Various global regulatory proteins including RpoS, OmpR, and CpxR were not involved in the regulation of the stf operon. A S. typhimurium $_X8661$ mutant lacking stfAC function attenuated 6.7 folds more than that of wild type $_X3761$ in the mouse virulence test, suggesting in the somehow involved in the Salmonella pathogenesis.

APPLICATION OF MULTIVARIATE DISCRIMINANT ANALYSIS FOR CLASSIFYING PROFICIENCY OF EQUIPMENT OPERATORS

  • Ruel R. Cabahug;Ruth Guinita-Cabahug;David J. Edwards
    • International conference on construction engineering and project management
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    • 2005.10a
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    • pp.662-666
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    • 2005
  • Using data gathered from expert opinion of plant and equipment professionals; this paper presents the key variables that may constitute a maintenance proficient plant operator. The Multivariate Discriminant Analysis (MDA) was applied to generate data and was tested for sensitivity analysis. Results showed that the MDA model was able to classify plant operators' proficiency at 94.10 percent accuracy and determined nine (9) key variables of a maintenance proficient plant operator. The key variables included: i) number of years of experience as equipment operator (PQ1); ii) eye-hand coordination (PQ9); iii) eye-hand-foot coordination (PQ10); iv) planning skills (TE16); v) pay/wage (MQ1); vi) work satisfaction (MQ4); vii) operator responsibilities as defined by management (MF1); viii) clear management policies (MF4); and ix) management pay scheme (MF5). The classification procedure of nine variables formed the general model with the equation viz: OMP (general) = 0.516PQ1 + 0.309PQ9 + 0.557PQ10 + 0.831TE16 + 0.8MQ1 + 0.0216MQ4 + 0.136MF1 + 0.28MF4 + 0.332MF5 - 4.387

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TOWARD A NEXT GENERATION SOLAR CORONAGRAPH: DEVELOPMENT OF A COMPACT DIAGNOSTIC CORONAGRAPH FOR THE ISS

  • Cho, K.S.;Bong, S.C.;Choi, S.;Yang, H.;Kim, J.;Baek, J.H.;Park, J.;Lim, E.K.;Kim, R.S.;Kim, S.;Kim, Y.H.;Park, Y.D.;Clarke, S.W.;Davila, J.M.;Gopalswamy, N.;Nakariakov, V.M.;Li, B.;Pinto, R.F.
    • Journal of The Korean Astronomical Society
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    • v.50 no.5
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    • pp.139-149
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    • 2017
  • The Korea Astronomy and Space Science Institute plans to develop a coronagraph in collaboration with National Aeronautics and Space Administration (NASA) and to install it on the International Space Station (ISS). The coronagraph is an externally occulted one-stage coronagraph with a field of view from 3 to 15 solar radii. The observation wavelength is approximately 400 nm, where strong Fraunhofer absorption lines from the photosphere experience thermal broadening and Doppler shift through scattering by coronal electrons. Photometric filter observations around this band enable the estimation of 2D electron temperature and electron velocity distribution in the corona. Together with a high time cadence (<12 min) of corona images used to determine the geometric and kinematic parameters of coronal mass ejections, the coronagraph will yield the spatial distribution of electron density by measuring the polarized brightness. For the purpose of technical demonstration, we intend to observe the total solar eclipse in August 2017 with the filter system and to perform a stratospheric balloon experiment in 2019 with the engineering model of the coronagraph. The coronagraph is planned to be installed on the ISS in 2021 for addressing a number of questions (e.g., coronal heating and solar wind acceleration) that are both fundamental and practically important in the physics of the solar corona and of the heliosphere.