• Title/Summary/Keyword: ompB

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Engineering of Recombinant Escherichia coli Towards Methanol Sensing Using Methylobacterium extroquens Two-component Systems

  • Selvamani, Vidhya;Ganesh, Irisappan;Chae, Sowon;Maruthamuthu, Murali kannan;Hong, Soon Ho
    • Microbiology and Biotechnology Letters
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    • v.48 no.1
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    • pp.24-31
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    • 2020
  • Five genes (mxbDM, mxcQE and mxaB) are responsible for the transcription of methanol oxidation genes in Methylobacterium strains. Among these, MxbDM and MxcQE constitute the two-component system (TCS) regulating methanol metabolism. In this study, we integrated the methanol-sensing domain of MxbD and MxcQ with the EnvZ/OmpR from Escherichia coli. The domain-swapping strategy resulted in chimeric histidine kinases (HK's) MxbDZ and MxcQZ AM1 containing recombinant E. coli. Real-time quantitative PCR was used to monitor OmpC expression mediated by the chimeric HK and response regulator (RR) OmpR. Further, an ompC promoter based fluorescent biosensor for sensing methanol was developed. GFP fluorescence was studied both qualitatively and quantitatively in response to environmental methanol. GFP measurement also confirmed ompC expression. Maximum fluorescence was observed at 0.05% methanol and 0.01% methanol using MxbDZ and MxcQZ AM1, respectively. Thus the chimeric HK containing E. coli were found to be highly sensitive to methanol, resulting in a rapid response making them an ideal sensor.

Antioxidant activity of Cinnamomum cassia extract and quality of raw chicken patties added with C. cassia powder and Pleurotus sajor-caju powder as functional ingredients during storage

  • Rakasivi, Kanita Galih Julia;Chin, Koo Bok
    • Animal Bioscience
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    • v.35 no.8
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    • pp.1279-1288
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    • 2022
  • Objective: The aim of this study was to investigate antioxidant activities of cinnamon (Cinnamomum cassia) extracts (extracted with different solvents) at various concentrations and to determine product quality of raw chicken patties added with different levels of cinnamon powder (CP) and oyster mushroon powder (OMP) during storage. Methods: After cinnamon was made into oven dried CP and extracted with water and different levels (50%, 80%, and 100%) of ethanol, antioxidant activities of these extracts were determined. CP and OMP were combined at different levels and added to raw chicken patties. Physicochemical properties and microbial counts were measured during refrigerated storage. Results: Cinnamon ethanol (80%) extract showed the highest (p<0.05) by 2,2-diphenyl-1picrylhydrazyl-radical scavenging activity and reducing power. Cinnamon water extract (CWE) had the highest iron chelating ability (p<0.05), while CP 100% ethanol extract had the highest content of total phenolic compound. Then, CP and OMP were applied to chicken patties at different levels (0.1% to 0.2%). After the addition of CPs, pH, L* (lightness), 2-thiobarbituric acid reactive substance, and volatile basic nitrogen values were decreased, whereas a* (redness) and b* (yellowness) values were increased. Microbial counts of total bacteria and Enterobacteriaceace were decreased with the addition of CP 0.2% regardless of the OMP level. Conclusion: The addition of CP in combination with OMP can increase the shelf-life of chicken patties during storage.

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.

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.

Occurrence of canine brucellosis in large kennels and characterization of Brucella canis isolates by PCR-RFLP (집단 개사육농장에서의 Canine Brucellosis 발생 및 PCR-RFLP를 이용한 분리주의 특성조사)

  • Kim, Jong-Wan;Lee, Young-Ju;Tak, Ryun-Bin
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.67-75
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    • 2003
  • A total of 260 dogs were randomly selected from two different treed kennels that brucellosis has occurred (group 1, 126 dogs), and random selected breed kennel (group 2, 134 dogs), and monitored for Brucella canis (B. canis) by 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) and bacterial culture method. For the differentiation, PCR-RFLP using omp-31, wbkA and per genes used for 52 of B canis strains (strain I) isolated in this study and 3 of B. canis strains (strain II) isolated in 1994 in Korea. 2ME-RSAT revealed that 63/126 dogs (50.0%) and 12/134 dogs (9.0%) were positive in group I and group II, respectively. Bacterial culture revealed that 47/126 dogs (37.3%) and 5/134 dogs (3.7%) were positive in group I and group II, respectively. As the results of PCR-RFLP, $\underline{omp}-31$ was amplified from all Brucella spp, except B. abortus. All B. canis isolates showed unique PCR-RFLP pattern following digestion with Bmel8I. However, all Brucella spp. showed the same PCR-RFLP pattern following digestion with SalI. PCR-RFLP analysis of wbkA revealed that all Brucella spp. showed the same pattern following digestion with HindIII. PCR-RFLP analysis of per revealed that B. abortus 544 and B. melitensis 63/9 showed the same pattern, but different from B. suis and B. canis following digestion with HindIII.

운용 및 유지보수 프로세서

  • 서장원;안규석;이규열
    • Information and Communications Magazine
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    • v.6 no.1
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    • pp.62-68
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    • 1989
  • TDX-1A의 유지보수 및 운용관리 프로세서는 8비트 프로세서 사용에 따른 성능상의 제약과 프로그램 체계상의 문제점이 있어 TDX-1B에서는 호처리에 직접 영향을 미치지 않는 SAP, SMP 및 IOIP를 32비트 프로세서로 통합하여 유지보수 및 운용관리를 체계화함과 동시에 이에 관련된 기능을 보완한 OMP(Operation and Maintenance Processor)를 개발하였다.

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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.

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.

A Novel GNSS Spoofing Detection Technique with Array Antenna-Based Multi-PRN Diversity

  • Lee, Young-Seok;Yeom, Jeong Seon;Noh, Jae Hee;Lee, Sang Jeong;Jung, Bang Chul
    • Journal of Positioning, Navigation, and Timing
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    • v.10 no.3
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    • pp.169-177
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    • 2021
  • In this paper, we propose a novel global navigation satellite system (GNSS) spoofing detection technique through an array antenna-based direction of arrival (DoA) estimation of satellite and spoofer. Specifically, we consider a sophisticated GNSS spoofing attack scenario where the spoofer can accurately mimic the multiple pseudo-random number (PRN) signals since the spoofer has its own GNSS receiver and knows the location of the target receiver in advance. The target GNSS receiver precisely estimates the DoA of all PRN signals using compressed sensing-based orthogonal matching pursuit (OMP) even with a small number of samples, and it performs spoofing detection from the DoA estimation results of all PRN signals. In addition, considering the initial situation of a sophisticated spoofing attack scenario, we designed the algorithm to have high spoofing detection performance regardless of the relative spoofing signal power. Therefore, we do not consider the assumption in which the power of the spoofing signal is about 3 dB greater than that of the authentic signal. Then, we introduce design parameters to get high true detection probability and low false alarm probability in tandem by considering the condition for the presence of signal sources and the proximity of the DoA between authentic signals. Through computer simulations, we compare the DoA estimation performance between the conventional signal direction estimation method and the OMP algorithm in few samples. Finally, we show in the sophisticated spoofing attack scenario that the proposed spoofing detection technique using OMP-based estimated DoA of all PRN signals outperforms the conventional spoofing detection scheme in terms of true detection and false alarm probability.

A Study on the Protein Productivity of the Promoters for Cold Inducible Genes in Escherichia coli (대장균 저온 유도성 유전자 Promoter의 단백질 생산성에 관한 연구)

  • Kim, So-Yeon;Kim, Su-Hyun;Heo, Mi-Ae;Lee, Sun-Gu
    • KSBB Journal
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    • v.21 no.6 s.101
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    • pp.461-465
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    • 2006
  • We investigated the protein productivity of the promoters for genes showing prolonged induction upon cold shock in Escherichia coli. Six low temperature inducible genes (frdA, glpB, hypB, katG, nupG, ompT) were selected based on the previously reported cDNA microarray based global transcription profiling of Escherichia coli Kl2 in response to cold shock. Their promoter regions were isolated from the genomic DNA of E. coli JM109 and expression levels induced by the promoters were examined by using green fluorescence protein (GFP) as a reporter at $15^{\circ}C$ and $37^{\circ}C$. Among the six promoters, the promoter for nupG showed the highest and prolonged expression at both temperatures and the cold inducibility of nupG promoter was not observed.