• 제목/요약/키워드: OmpC

검색결과 54건 처리시간 0.03초

TaqMan 실시간 중합 효소 연쇄반응에 의한 살모넬라속의 검출 및 ompC 항원단백 유전자의 비교 (Detection of Salmonella spp. by TaqMan real-time PCR and comparison of nucleotide sequences of ompC gene among Salmonella)

  • 이영성;최경성;김명철;한재철;채준석
    • 대한수의학회지
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    • 제42권4호
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    • pp.513-522
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    • 2002
  • Antigenic ompC genes of S. gallinarum, S. pullorum and S. dublin were characterized among Salmonella spp. isolated from chickens and other animals to identify genetic variation. Salmonella ompC gene fragment (1,027 bp) was amplified by PCR and the amplicons were cloned for comparison of nucleotide sequences. The identity of the sequences between S. gallinarum and S. pullorum, S. gallinarum and S. dublin, S. pullorum and S. dublin was 99.8%, 97.6% and 97.8%, respectively. Also, we found that ompC has some diversity between S. gallinarum and S. pullorum, and other Salmonella spp. which may be useful to type the organisms. Similar to diagnosis in other organisms, the TaqMan PCR method can be applied to rapid and accurate diagnosis of salmonellosis in chickens and other animals. We designed PCR primers and TaqMan probe for flagellin gene (fliC) for detection of Salmonella spp. by TaqMan PCR. The TaqMan PCR method was 10,000 times more sensitive than conventional PCR.

Effect of Cimetidine and Phenobarbital on Metabolite Kinetics of Omeprazole in Rats

  • Park Eun-Ja;Cho Hea-Young;Lee Yong-Bok
    • Archives of Pharmacal Research
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    • 제28권10호
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    • pp.1196-1202
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    • 2005
  • Omeprazole (OMP) is a proton pump inhibitor used as an oral treatment for acid-related gastrointestinal disorders. In the liver, it is primarily metabolized by cytochrome P-450 (CYP450) isoenzymes such as CYP2C19 and CYP3A4. 5-Hyroxyomeprazole (5-OHOMP) and omeprazole sulfone (OMP-SFN) are the two major metabolites of OMP in human. Cimetidine (CMT) inhibits the breakdown of drugs metabolized by CYP450 and reduces, the clearance of coad-ministered drug resulted from both the CMT binding to CYP450 and the decreased hepatic blood flow due to CMT. Phenobarbital (PB) induces drug metabolism in laboratory animals and human. PB induction mainly involves mammalian CYP forms in gene families 2B and 3A. PB has been widely used as a prototype inducer for biochemical investigations of drug metabolism and the enzymes catalyzing this metabolism, as well as for genetic, pharmacological, and toxicological investigations. In order to investigate the influence of CMT and PB on the metabolite kinetics of OMP, we intravenously administered OMP (30 mg/kg) to rats intraperitoneally pretreated with normal saline (5 mL/kg), CMT (100 mg/kg) or PB (75 mg/kg) once a day for four days, and compared the pharmacokinetic parameters of OMP. The systemic clearance ($CL_{t}$) of OMP was significantly (p<0.05) decreased in CMT-pretreated rats and significantly (p<0.05) increased in PB-pretreated rats. These results indicate that CMT inhibits the OMP metabolism due to both decreased hepatic blood flow and inhibited enzyme activity of CYP2C19 and 3A4 and that PB increases the OMP metabolism due to stimulation of the liver blood flow and/or bile flow, due not to induction of the enzyme activity of CYP3A4.

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

  • 김명석;진지웅;정승희;서정수;홍수희
    • 수산해양교육연구
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    • 제27권5호
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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.

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
    • 한국미생물·생명공학회지
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    • 제48권1호
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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.

A Brucella Omp16 Conditional Deletion Strain Is Attenuated in BALB/c Mice

  • Zhi, Feijie;Fang, Jiaoyang;Zheng, Weifang;Li, Junmei;Zhang, Guangdong;Zhou, Dong;Jin, Yaping;Wang, Aihua
    • Journal of Microbiology and Biotechnology
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    • 제32권1호
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    • pp.6-14
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    • 2022
  • Brucella spp. are facultative intracellular pathogens that invade, survive and proliferate in numerous phagocytic and non-phagocytic cell types, thereby leading to human and animal brucellosis. Outer membrane proteins (Omps) are major immunogenic and protective antigens that are implicated in Brucella virulence. A strain deleted of the omp16 gene has not been obtained which suggests that the Omp16 protein is vital for Brucella survival. Nevertheless, we previously constructed an omp16 conditional deletion strain of Brucella, ∆Omp16. Here, the virulence and immune response elicted by this strain were assessed in a mouse model of infection. Splenomegaly was significantly reduced at two weeks post-infection in ∆Omp16-infected mice compared to infection with the parental strain. The bacterial load in the spleen also was significantly decreased at this post-infection time point in ∆Omp16-infected mice. Histopathological changes in the spleen were observed via hematoxylin-eosin staining and microscopic examination which showed that infection with the ∆Omp16 strain alleviated spleen histopathological alterations compared to mice infected with the parental strain. Moreover, the levels of humoral and cellular immunity were similar in both ∆Omp16-infected mice and parental strain-infected mice. The results overall show that the virulence of ∆Omp16 is attenuated markedly, but that the immune responses mediated by the deletion and parental strains in mice are indistinguishable. The data provide important insights that illuminate the pathogenic strategies adopted by Brucella.

오메프라졸과 메글루민의 복합체 형성과 안정성 (Complexation of Omeprazole with Meglumine and its Stability)

  • 이계주;김성욱;도기찬;박종범;황성주
    • Journal of Pharmaceutical Investigation
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    • 제27권4호
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    • pp.253-263
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    • 1997
  • To investigate the interaction of omeprazole (OMP) and meglumine (MEG), a complex was prepared by freeze-drying method in ammoniacal aqueous medium at room temperature and subjected to IR, DSC, and 1H NMR analysis. In addition, the stability of the complex was tested by accelerated stability analysis, and the dissolution rate of both powder and enteric coated was determined pellet by paddle method. The results are as follows; i) IR, DSC, and $^{1}H$ NMR studies indicate the formation of inclusion complex between OMP and MEG probably by electrostatic forces as $[OMP]\;[MEGH]^+$ form in a stoichiometric ratio (1:1) of OMP : MEG. ii) The dissolution rate of enteric coated OMP-MEG complex pellet in simulated enteric fluid was 90.6% in 10 minutes, which may satisfy the requirement for the regulation of dissolution. iii) OMP-MEG complex were decomposed according to pseudo 1st order kinetics: while the decomposition of OMP showed a rate constant $(k_{25^{\circ}C})$ of $5.13{\times}10^{-4}{\cdot}\;day^{-1}$, a half-life$(t_{1/2})$ of 1,350 days, a shelf-life$(T_{90%})$ 205 days and an activation energy of 23.53 kcal/mole. OMP-MEG complex inhibited a rate $(k_{25})$ of $2.92{\times}10^{-4}{\cdot}\;day^{-1}$, a half-life$(t_{1/2})$ of 2,373 days, a shelf-life $(T_{90%})$ of 306 days and an activation energy of 20.18 kcal/mole. iv) OMP was stabilized markedly by the formation of OMP-MEG complex between OMP and MEG, and the humidity increased the stability of OMP-MEG complex by decreasing the decomposition rate$(k_{50^{\circ}C})$ from $1.27{\times}10^{-2}{\cdot}\;day^{-1}$ at 31% R.H. to $2.54{\times}10^{-2}{\cdot}\;day^{-1}$ at 90% R.H.

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참외과피추출물을 적용한 양송이의 저장 중 품질 변화 (Use of oriental melon peel extracts to maintain the quality of Agaricus bisporus during its storage)

  • 박혜진;김건희
    • 한국식품저장유통학회지
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    • 제21권4호
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    • pp.473-482
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    • 2014
  • 참외과피추출물의 천연갈변저해제로서의 가능성을 확인하기 위하여 참외과피추출물 및 ascorbic acid를 적용한 양송이의 저장 중 품질변화를 확인하였다. 양송이를 각 처리구별 PP film으로 포장하여 4 및 $15^{\circ}C$에서 저장하면서 포장 내 기체조성, 경도, 표면 색 및 관능평가를 관찰하였다. 그 결과 기체조성 또는 경도의 변화에 있어서는 처리구별 큰 차이 없이 저장온도 또는 포장재에 따른 영향을 받았으나, 갈변저해효과에 있어서는 참외과피추출물을 처리하였을 때 양성대조군으로 사용한 ascorbic acid 보다 표면 색 유지에 효과적이었다. 또한 관능평가에서도 참외과피추출물을 적용한 처리구(OMP, OMP+AA)에서 저장기간 동안 높은 점수를 유지하였다. 참외과피에 함유되어있는 총 페놀과 플라보노이드의 함량을 조사한 결과, 참외과피추출물의 총 페놀함량은 플라보노이드 함량보다 4배 정도 높게 나타났다. 참외과피추출물의 항산화 및 항갈변 활성은 DPPH 라디칼 소거활성과 PPO 저해활성이 낮은 농도에서도 높은 활성을 나타냈으나 ABTS 라디칼 소거활성과 구리킬레이트 활성에서는 비교적 높은 농도에서 활성을 보였다. 이에 따라 참외과피추출물을 천연갈변저해제로서 양송이에 적용하였을 때 효과적으로 양송이의 갈변을 저해할 수 있음을 확인하였다.

Surface Display of $Poly(His)_n$ on the Escherichia coli using OmpC as Surface Anchoring Motif

  • 최종현;;이상엽
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 춘계학술발표대회
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    • pp.523-526
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    • 2000
  • A novel cell surface display system was developed by employing the Escherichia coli outer membrane protein C (OmpC) as an anchoring motif. Poly-histidine (poly-His) peptides of 19, 32, 45, 84, and 162 amino acids (aa) could be successfully displayed by inserting them into the seventh exposed loop(L7) of OmpC. Recombinant cells displaying poly-His of 19, 32, 45, and 84 aa could absorb 18.9, 23.9, 26.1, and 32.0 ${\mu}mol$ of $Cd^{2+}$ per gram cell dry weight, respectively and therefore, would be useful as the biosorbents of heavy metals.

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열 및 에탄을 shock이 Vibrio vulnificus의 막단백질에 미치는 영향 (Effects of heat and ethanol shock on the membrane proteins of Vibrio vulnificus)

  • 허문수;정초록
    • 한국어병학회지
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    • 제12권2호
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    • pp.89-99
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    • 1999
  • V. vulnificus ATCC 27562균주의 배양 온도를 $2^{\circ}C $, 20분간 및 6% 에탄올, 10분간으로 반응시켰을 때 SDS-PAGE상에서 새로운 16가지의 heat shock protein(hsps)과 10가지의 ethanol shock protein이 나타났다. Lethal temperature에 노출하기전에 미리 열 충격을 가한 경우 thermo tolerance가 유도되었다. 균체면역에 의해 생성된 항혈청과 열 충격 세포에서 분리된 막단백질과의 ELISA에서는 Outer Membrane Protein(OMP)에서 높은 면역반응을 나타냈으며 western blotting으로는 Inner Membrane Protein(IMP)에서는 62kDa, OMP에서는 69 kDa단백이 높은 면역원성을 나타냈다. ethanol 충격 반응에서는 IMP에서는 48 kDa, OMP에서는 오직 major밴드에서만 면역반응성이 확인되었다. anti-V, vulnificus혈청에 대한 균체 응집시험에서는 열 충격 반응 후의 균체가 정상 균체에 비해 응집반응성이 높았다.

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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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    • 제35권8호
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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.