• 한국해양바이오학회지
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• 제2권2호
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• pp.123-125
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• 2007

Vibrio anguillarum is a gram-negative bacterium that causes vibriosis in approximately 80 different fish species. V. anguillarum produces several exotoxins are correlated with the pathogenesis of vibriosis. This study is focused on the composition of the outer membrane vesicle. Most of gram-negative bacteria produce outer membrane vesicle (OMV) during cell growth. OMV was formed from the outer membrane surface of cell and than released to extracellular environment. OMV consists of outer membrane lipids, outer membrane protein (OMP), LPS, and soluble periplasmic components. Also, they contain toxins, adhesions, and immunomodulatory. Many gram-negative bacteria were studied out forming OMV. In Vibrio sp., formation of OMV by electron microscopy has been reported from V. cholerae and V. parahaemolyticus. In present study, we isolated OMV from V. anguillarum and OMV protein was separated by SDS-PAGE. Magor band was sliced and analyzed by MALDI-TOF. The major protein band of 38kDa was identified as OmpU by MALDI-TOF MS analysis.

• Journal of Microbiology
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• 제43권5호
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• pp.437-442
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• 2005

Sera from rabbits were infected with Vibrio vulnificus containing an antibody against major outer membrane protein (MOMP). MOMP of V. vulnificus ATCC 27562 were isolated and purified by Sarkosyl and TritonX-100 dual treatment. Molecular size of MOMP was identified as 36-kDa on $13\%$ SDS-PAGE. The sequence of the first 26 amino acid residues from the N-terminal end of the protein is AELYNQDGTSLDMGGRAEARLSMKDG, which is a perfect match with OmpU of V. vulnificus CMCP6 and YJ016. MOMP specific IgM and IgG were investigated in groups of mice. The group of mice immunized with MOMP and Alum showed higher levels of IgG2b than the group immunized with only MOMP. Vaccination with MOMP resulted in protective antibodies in the mouse infection experiment.

• 대한수의학회지
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• 제46권1호
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• pp.13-19
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• 2006

Generally known psittacosis or ornithosis is a disease of birds caused by the bacterium Chlamydia psittaci. Humans are accidential hosts and are most commonly infected from avian sources. It raises hepatitis or neurosis. As major outer membrane protein (MOMP) of Chlamydia psittaci has been known to play a role in the avoidance of host immune defenses, research on developing a Chlamydia vaccine has focused on the MOMP. In this study, the gene encoding the major outer membrane protein (MOMP) of the Chlamydia psittaci strain 6BC was cloned and expressed in Escherichia coli strain M-15. The recombinant DNA was cloned by fusion prokaryotic expression vector pQE30-GFPII. Expression of the recombinant protein was performed in E. coli and was induced by IPTG. The size of expressed recombinant protein is 74.220 kDa (MOMP, 43.260 kDa; GFP expression region, 30 kDa; $6{\times}His$ tag, 960Da). This protein was purified by using his-tagging-inclusion body. Recombinant protein was reconfirmed through ELISA test and western blot with antibody against pQE30-GFPII. It will be useful antibody development.

• Journal of Life Science
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• 제9권1호
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• pp.35-39
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• 1999

Elecrophoreticl analysis of a 36 kDa protein was runned by SDS-PAGE, isoelectric focusing (IEF) and two dimensional electrophoresis pattern. Major 36 kDa and 25, 46, 48, 66 kDa protein were detected by Coomassie blue stain on SDS-PAGE. Major 36kDa protein was eluted for production of antiserum for serological analysis, IEF and two dimensional electrophoresis. Isoelectric point of 36kDa was aout pH 8.5. Two dimensional electrophoresis of eluted 36kDa showed one point on the gel. Anti-36 kDa serum made by newzilland rabbit for serological test. In ELISA, final titer of antibody was 100×{TEX}$2^5}${/TEX} : 1. Neutralize ability of serum was examined by slide agglutination test and colonization test in rat. Anti-36 kDa serum agglutinated whole cell of V. vulnificus were inhibited colonization on intestine in rat. Accordingly In this paper contain some electrophoretical analysis and serological test of a 36 kDa OMP of V. vulnificus.

• Journal of Photoscience
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• 제9권2호
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• pp.47-50
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• 2002

Raft is a distinctive membrane domain enriched in a certain class of lipids, cholesterol, and proteins observed on the plasma membrane. Growing evidence has revealed that such membrane domains play key roles in signal transduction, fertilization, development, transmitter release, and so on. Recently, we have isolated raft-like detergent-resistant membrane (DRM) fraction from bovine photoreceptor rod outer segments. Transducin and its effecter, cGMP-phosphodiesterase, elicited stimulus-dependent translocation between detergent-soluble membrane and DRM. This suggested potential importance of such distinct membrane domains in vertebrate phototransduction. Here, we will discuss physiological meaning of the translocation of major components of cGMP cascade to raft-like membrane in phototransduction. We would like to propose a hypothesis that raft-like membrane domains on the disk membrane are the place where cGMP cascade system could be quenched.

• 한국환경독성학회:학술대회논문집
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• 한국환경독성학회 2001년도 춘계심포지움 및 학술발표회
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• pp.140-140
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• 2001

Porins are major outer membrane proteins which produce non-specific aqueous channels across the membrane that permit the diffusion into the bacterial cells of hydrophilic compounds including sugars, amino acids, and antibiotics. In some gram-negative organisms, antibiotic resistance can be induced by mutational loss of channel that causes a decrease in outer membrane permeability. (omitted)

• 미생물학회지
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• 제43권1호
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• pp.7-13
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• 2007

Pasteurella multocida는 돼지에서 위축성 비염, 폐렴을 비롯한 다양한 호흡기 질환을 일으키는 병원균이다. 본 연구에서는 돼지 위축성 비염에 대한 효과적인 순수정제 백신을 개발하고자 하는 기초 연구로서 P. multocida의 외막 단백질 H에 의해 유도되는 방어적 항체와 면역을 확인하였다. P. multocida의 외막 단백질을 포함하는 분획은 호흡기 질병 혼합 백신에 대한 항혈청과 불활화된 사균 세포에 대한 항혈청 모두에서 면역학적으로 검출 가능하였다. 선행 연구에서 분리한 외막 단백질 H 유전자는 재조합 발현 백터 제작에 이용되어 대장균으로부터 재조합 외막단백질 H를 정제하였다. 실험 동물 면역과 항혈청의 교차반응, ELISA를 통한 항체 역가의 측정 및 공격접종을 통하여, 재조합 외막 단백질 H는 높은항원성을 가지며, 지속적인 체액성 면역을 유도하는 것을 확인하였다. 외막 단백질 H는 순수정제 항원으로서 P. multocida에 의한 호흡기 질환에 대한 효과적인 방어를 유도할 수 있는 단위 백신 후보 단백질로 여겨진다.

• 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.

• 한국미생물·생명공학회지
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• 제25권2호
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• pp.129-136
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• 1997

Helicobacter pylori is a spiral-shaped, microaerophilic human gastric pathogen causing chronic-active gastritis in association with duodenal ulcer and gastric cancer. To investigate the possibility of H. pylori outer membrane proteins (OMPS) as the oral vaccine antigens, sarcosine-insoluble outer membrane fraction has been prepared from H. pylori NCTC 11637. The major OMPs having apparent molecular masses of 62 kDa, 54 kDa and 33 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which were identified as urease B subunit (UreB), heat shock protein (Hsp54 kDa) and urease A subunit (UreA), respectively. Minor protein bands of 57 kDa, 52 kDa, 40 kDa, 36 kDa and 31 kDa were also observed. The antigenicity of H. pylori OMPs and antigenic cross-reactivity among the strains were determined by immunoblot analysis using anti-H. pylori OMPs antisera or intestinal lavage solutions. The results showed that UreB, Hsp54 kDa, UreA and 40 kDa proteins vigorously stimulated mucosal immune response rather than systemic immunity. From this results, these proteins seemed to be useful as the antigen candidates for the oral vaccine. The immunoblotting results with surface proteins from eight isolated H. pylori strains were similar to that of H. pylori NCTC 11637. The IgA which had been arised from oral administration of H. pylori OMPs, was able to bind H. pylori whole-cells.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2005년도 제45회 학술심포지움 및 추계학술대회
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• pp.83-83
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• 2005