• 한국잠사학회:학술대회논문집
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• 한국잠사학회 2003년도 제46회 춘계 학술연구 발표회
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• pp.43-43
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• 2003

잠란의 휴면 개시, 유지 및 각성 등에 따른 단백질의 변화를 알아보기 위해 2D-전기영동에 의한 peptides 분석을 하였다. 잠품종은 백옥잠을 사용하였으며, 단백질 분석은 산란 후 5일 경과란(휴면란), 산란 후 20시간에 침산하여 1일 및 2일 경과란, 및 산란 후 2일 경과 후 냉장(5$^{\circ}C$)처리하여 1일, 3일, 5일된 잠란을 각각 대상으로 하였다. 2D-전기영동은 1차로 pH3-10 range에서 isoelectric focusing 하고 2차로 SDS-PAGE한 후 silve stainin으로 peptides를 검출하였다. (중략)

• Restorative Dentistry and Endodontics
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• 제20권1호
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• pp.71-95
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• 1995

Bacteria have been regarded as a one of major etiologic factors in root canal infections. In endodontic treatment the effective removal of pathogenic microorganisms in the root canal is the key to successful outcome. Bacterial cell wall components may play an important role in the development of pulpal and periapical disease. The purpose of this study was to evaluate the effect of sonic extracts of Streptococcus spp. on cultured L929 cells and to characterize cell wall protein profiles of Streptococcus spp. Streptococcus spp. were isolated from infected root canals and identified with Vitek Systems(Biomeriux, USA). Five streptococci, namely S. sanguis, S. mitis, S uberis, S. mutans (ATCC 10449) and S. faecalis (ATCC 19433) weere enriched in brain heart infusion broth. Cell pellets were sonicated and cell wall extracts were dialyzed and membrane filtered. Prepared cell wall proteins were applied to cultured L929 cell. The cell reaction were evaluated by monitoring DNA synthesis, cell numbers and the change of cell morphology. The total cell wall protein profiles of microorganisms were characterized by sodium dodecyl sulfate polyacrylamide-gel eledruphoresis(SDS-PAGE). DNA synthesis of L929 cells were reduced by the increasing concentration of sonic extracts. DNA synthesis was significantly suppressed in more than $50{\mu}g$/ml of sonic extract conentration in five streptococci. S. nutans (ATCC 10449) showed stronger suppression on DNA synthesis than remaining four streptococci, which had the similar effect on DNA synthesis. Analysis of DNA synthesis measured by [$^3H$]-thymidine uptake was more sensitvie method than cell counting. Sonic extracts affected the microscopic findings of L929 cells. The protein profiles indicated that all five strains shared two major proteins with molecular masses of 70.8 and 57.5 kD respectively. S. uberis and S. mutans shared common minor proteins of which molecular weights were 147.9 and 112.2 kD respectively. However some minor proteins were unique for S. mitis, S. uberis and S. faecalis.

• 한국잡초학회지
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• 제16권4호
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• pp.354-361
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• 1996

Thiobencarb 처리에 의한 수도품종의 내성 정도를 조사하고 내성 메카니즘을 연구하기 위하여 내성 및 감수성 품종간의 단백질 함량, SDS-PAGE에 의한 단백질 유형의 변화를 조사하였다. 본 시험에 공시한 5개 품종 중 IR 10198-66-2와 IR 9660-50-3-1은 온실 및 실내실험에서 공히 thiobencarb에 대하여 내성을 보였으며 IR 22, IR 31802-48-2-2, IR 20656-R-R-R-6-1은 제초제 농도가 증가함에 따라 생장이 크게 억제되어 감수성을 나타내었다. Thiobencarb 처리에 의한 단백질의 함량은 내성 및 감수성 품종간에 뚜렷한 차이를 보였는데 감수성 품종의 단백질 함량은 제초제 농도가 증가함에 따라 크게 감소하는 경향을 나타내었으나 내성품종의 경우 감소가 거의 없었다. SDS-PAGE에 의한 단백질 밴드는 내성 품종의 경우 5ppm 처리에서도 변화가 없었으나 감수성 품종은 5ppm 처리에서 94-30 kD 사이와 14.4kD 부근의 단백질 밴드가 사라지는 경향을 보였다. Thiobencarb 3ppm을 처리한 감수성 품종인 IR 22의 2차원 전기영동에 의한 단백질 패턴에서는 14.4kD와 55kD 부근의 spot가 사라지거나 density가 감소하는 경향을 보였다. 이상의 단백질 함량과 유형의 변화로 미루어 보아 thiobencarb 처리는 감수성 품종의 특정 단백질 합성을 억제하는 것으로 나타났으며, 벼의 thiobencarb에 대한 내성은 이 부위의 단백질과 밀접한 관계가 있는 것으로 사료된다.

• Journal of Ginseng Research
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• 제13권2호
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• pp.158-164
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• 1989

인삼엽소병(leaf-burning disease) 원인과 light-harvesting chlorophyll-protein(LHCP) complex의 solar energy 분배능력과의 상호 연관성을 조사하기 위한 기초 연구로써 인삼 thylakoid의 chlorophyll-protein(CP) complex의 조성 및 특징을 조사하였다. 인삼의 CP-complex는 non-denaturing SDS-PAGE 방법에 의해 4개 bands로 분리되었으며 각 band는 Bassi와 Dunahay의 결과에 따라 CPI(PSI의 reaction center와 LHCP I antennae), CP I(PSI reaction center), LHCP II(LHCP II)의 oligoform), 그리고 LHCP II(PS II antennae; CP29, CP26)로 확인되었다. 인삼의 LHCP II 는 양지식물인 spinach, soybean과 비교해 볼 때 오히려 인삼의 band intensity가 더 높았으며, CP I band는 인삼에서만 분리되었다. 인삼 CP-complex band의 absorption 및 fluorescence spectra, chlorophyll a.b ratio 에서도 비교식물과 차이를 나타내었다. Thylakoid membrane의 polypeptide 함량은 인삼에서 비교식물에 비해 현저히 낮은 polypeptide 함량은을 나타내었다. SDS-PAGE에 의한 polypeptide pattern은 band의 수나 band intensity에서 비교식물과 차이를 나타내었으며, 특히 29-35 kD, 55 kD과 60 kD 근치에서 현저한 band intensity 차이를 확인하였다. Specific $^1O_2$에 의해 chl. a가 60%, chl.b는 90%, 그리고 carotenoid는 70%가 파괴되는 것으로 확인되었다.

• Advances in nano research
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• 제6권1호
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• pp.81-92
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• 2018

Biosynthesis of nanoparticles has acquired particular attention due to its economic feasibility, low toxicity and simplicity of the process. Extracellular synthesis of nanoparticles by bacteria and fungi has been stated to be brought about by enzymes and other reducing agents that may be secreted in the culture medium. The present study was carried out to determine the underlying mechanisms of extracellular silver nanoparticle synthesis by Pseudomonas hibiscicola isolated from the effluent of an electroplating industry in Mumbai. Synthesized nanoparticles were characterized by spectroscopy and electron microscopic techniques. Protein profiling studies were done using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (1D-SDS PAGE) and subjected to identification by Mass Spectrometry. Characterization studies revealed synthesis of 50 nm nanoparticles of well-defined morphology. Total protein content and SDS PAGE analysis revealed a reduction of total protein content in test (nanoparticles solution) samples when compared to controls (broth supernatant). 45.45% of the proteins involved in the process of nanoparticle synthesis were identified to be oxidoreductases and are thought to be involved in either reduction of metal ions or capping of synthesized nanoparticles.

• 한국동물학회지
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• 제38권4호
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• pp.490-497
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• 1995

미국흰불나방 의지방체 조직을 [35S]-메타이오닌이 포하된 배지에서 조직배양한 결과, 저장단백질-1(SP-1)의 전용기부터 용 1일 사이에 지방체로 흡수됨을 알았다. CHAPS, Triton X-100 등의 계면활성제를 농도별로 처리하여 막단백질의 용해도를 스크리닝한 뒤, anti-SP-1 polyclonal 항체를 쓴 Western blotting과 ligand blotting, 그리고 in vitro reductive methylation으로 14C을 표지한 저장단백질-1을 사용한 fluorography 등으로 1개의 수용체 밴드를 확인하였다. 1% Triton X-100으로 용해시킨 부분정제하였고, SDS-PAGE에 의해서 분자량을 측정한 결과 약 80 kDa로 나타났고 isoelectric focusing 시행 결과 등전점은 약 6.1로 계산되었다. 수용체 분자는 환원조건과 비환원조건의 차이와 전기영동 중의 온도에 따라서 SDS-PAGE상의 뚜렷한 밴드 양상의 차이를 나타내었다.

• BMB Reports
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• 제39권1호
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• pp.22-25
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• 2006

The gene encoding MPB83 from Mycobacterium bovis Vallee111 chromosomal DNA was amplified by using polymerase chain reaction (PCR) technique, and the PCR product was approximately 600bp DNA segment. Using T-A cloning technique, the PCR product was cloned into pGEM-T vector and the cloning plasmid pGEM-T-83 was constructed successfully. pGEM-T-83 and pET28a(+) were digested by BamHI and EcoRI double enzymes. The purified MPB83 gene was subcloned into the expression vector pET28a(+), and the prokaryotic expression vector pET28a-83 was constructed. Plasmid containing pET28a-83 was transformed into competence Escherichia coli BL21 (DE3). The bacterium was induced by isopropyl-$\beta$-D-thiogalactopyranoside (IPTG) and its lysates were loaded directly onto sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), approximately 26 kDa exogenous protein was observed on the SDS-PAGE. The protein was analyzed using Western-blotting. The results indicated that the protein was of antigenic activity of M. bovis. The results were expected to lay foundation for further studies on the subunit vaccine and DNA vaccine of MPB83 gene in their prevention against bovine tuberculosis.

• International Journal of Industrial Entomology and Biomaterials
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• 제23권1호
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• pp.147-153
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• 2011

The Pseudorabies (PR), also called Aujeszky's disease (AD), is an infectious viral disease caused by an alpha herpes virus and has domestic and wild pigs, as well as a wide range of domestic and wild animals, as the natural host. Pseudorabies virus (PRV) virions contain several envelope glycoproteins. Among them, gB, gC and gD are regarded as the major immunogenic proteins. We expressed these glycoproteins using the bacterial expression system and analyzed recombinant proteins. Expression of glycoproteins gC and gD were observed on SDS-PAGE or Western blot analysis, but gB was not. Optimal concentration of IPTG and inducing time were determined as 1.0 mM and 4 h, respectively, for the expression of both gC and gD in E. coli. A sodium dodecyl sulfate (SDS) was the most efficient detergent in solubilizing insoluble recombinant protein.

• 대한한방소아과학회지
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• 제13권1호
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• pp.253-275
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• 1999

The effects of Sagunjatang and Samultang on the immunosuppression induced by methotrexate(MTX) in rats were investigated in this study. The multiple parameters of immunity assessed in each rats included leukocyte count, lymphocyte rate, the number of lymphocyte in tibial bone marrow, contact hypersensitivity to DNFB, morphological change of thymocyte and IgG antibody on SDS-PAGE. Sprague-Dawley male rats were used and divided into five groups at random. Group A was normal control. Group B, the MTX treatment control, was injected i.v. with 2mg/kg of on days 9, 11 after sensitization with SRBC on 5th day. Group C, the experimental control, was treated Sagunjatang for 18days and MTX. Group D was treated Samultang for 18days and MTX. Group E was treated Sagunjatang-Samultang for 18days and MTX. The dosage of Sagunjatang and Samultang was $1m{\ell}/day$ respectively. In the case of Group E, rats Were fed Sagunjatang $1m{\ell}$ in the morning and Samultang $1m{\ell}$ in the afternoon. The results are summarized as follows: 1. Leukocyte count in rats induced by intravenous sensitization with SRBC was decreased significantly in Group E. 2. Leukocyte counts of 2weeks later after being treated MTX were increased significantly in Groups C and D. 3. Lymphocyte rate in rats induced by intravenous sensitization with SRBC wasn't changed significantly in all the experimental groups. 4. Lymphocyte rate of 2weeks. later after being treated MTX was increased significantly in Group D. 5. The number of lymphocyte in tibial bone marrow was incereased significantly in Group C. 6. Contact hypersensitivity wasn't changed significantly in all the experimental groups. 7. Morphological finding of thymocyte in group C was similar to normal group as compared with control group. 8. Purified IgG of all the experimental groups showed two bands of 50,000 and 25,000 on SDS-PAGE. But there was no difference among experimental groups.

• Parasites, Hosts and Diseases
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• 제26권2호
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• pp.73-86
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• 1988

간흡충(Clonorchis sinensis)의 조항원(WWA), 정제항원(ABA)의 민감도와 특이도를 비교해 보는 한편, 발육단계별로 충란 항원(EGA)-피낭유충 항원(MEA)-성충 항원(WWA)의 단백질 구성물질의 차이를 SDS-polyacrylamide gel electrophoresis(SDS-PAGE)로 관찰하고 이들 항원의 유용성을 효소면역반응(enzyme-linked immunosorbent assay: ELISA)으로 비교하여 다음과 같은 결과를 얻었다. 간흡충의 조항원으로부터 정제된 항원(ABA)은 항체와 결합한 결합항원(affinity-purified antibody(IgG) binding antigen:ABA)으로서 Ouchterlony 반응으로 간흡충 감염 가토혈청과 4개의 침모대 (WWA는 7개)를 형성하였다. WWA의 major protein들은 분자량 16,300∼18,500 및 28,000∼29,000 dalton의 Polypeptide band였고, ABA는 분자량 18,000∼21,000 및 29,000∼31,000 dalton의 단백질로 다소의 차이를 보였다. ELISA시첩에서 ABA는 WWA에 비해 현저히 민감도가 낮았다. WWA와 ABA에 대한 폐흡충 감염 사람혈청의 ELISA 정사에서 WWA는 Ouchterlony 양성자 8예중 3예에서 교차반응을 나타내었으나 ABA는 교차반응을 나타내지 않았다. 간흡충의 발육단계별 항원 단백질의 분자량 범위는 WWA 11,000∼80,000, MEA 15,000∼100,000, EGA 15,000∼200,000 dalton이었다. 주류원 단백질의 분자량은 EGA는 각각 36,600 (band No. 22), 38,500(No. 20), 64,000(No. 9), 62,000(No. 10), 54,500(No. 11), 53,000(No. 12) dalton, MEA는 각각 65,600(No. 3), 44,700 (No.7), 43,900(No. 8) dalton, WWA는 각각 16,300∼18,500(No. 31-32), 28,000∼29,000(No. 25), 11,000w13,000 (No. 35) 및 31,000(No. 24) dalton이었다. 즉, MEA와 EGA가 WWA보다 고분자의 단백질로 구성되어 있었다. 각 발육단계별 항원의 항원성은 ELISA 반응으로 볼 때 WWA가 가장 높았고, MEA는 약한 항원성만을 나타내었으며 EGA는 음성이었다. WWA와 간흡충 감염 가토혈청은 감염 4∼6주에 OD>1.0, 12주 이후 항체가의 plateau를 나타내었으나 MEA는 Ouchterlony 반응에서 EGA와 함께 음성반응을 보였다. ELISA에서도 중감염군(12∼20주)에서만 OD>0.6으로 미약한 항원성이 인정되었으며 경감염군은 전감염기간중 OD<0.6를 나타내었다. 이상의 결과로 보아 성충 항원이 가장 강력한 함원성 물질을 포함하고 있음이 명백하다고 생각된다. 그러나 성충 조항원은 면역진단에 사용할 겹우 타흡충과의 빈번한 교차반응이 예상되므로 민감도 및 특이도가 높은 정제 항원물질을 분리하는 일이 중요하다고 생각된다.