• 한국미생물·생명공학회지
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• 제23권1호
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• pp.110-117
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• 1995

This study was carried out to immunologically characterize Bacillus thuringiensis (B.t) antigens. Protein patterns of ultrasonicated- antigens of B. thuringiensis subspecies using SDS- PAGE revealed marked similarities among all the strains analyzed except for the difference between quantative variations of bands and some protein antigens. The comparison of the protein patterns showed that the protein antigen of 45 kilodalton (kd) was common in 11 strains and that the difference between B. thuringiensis subsp. canadensis and galleriae was noticed in quantative variations of bands despite of ambiguous serogrouping, suggesting a useful method for identification. All strains examined showed similar antigenic patterns in SDS-PAGE, while immunodominant bands differed in antigenic reactivity in western blot using polyclonal antibodies. Polyclonal antibody to B. thuringiensis subsp. thuringiensis and israelensis in indirect immunofluorescence assay reacted with flagella and cell surface antigens. The present study indicates that SDS-PAGE and western blot analysis may be used as tools for differentiation and identification of B. thuringiensis subspecies.

• The Plant Pathology Journal
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• 제18권3호
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• pp.126-129
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• 2002

Infected sweet potato (Ipomoea batatas) showing symptoms of sunken veins, stunting, mosaic, and mottling were collected from Gimje, Cochang, Iksan, and Haenam provinces in Korea. Electron microscopic (EM) observation of the infected tissue revealed rod and filamentous rod type virus particles of various lengths. Western blot analysis of the protein samples extracted from infected sweet potato and partially purified virus identified the isolates as Sweet potato feathery motile virus (SPFMV), Sweet potato latent virus (SwPLV), and Sweet potato chlorotic stint virus (SPCSV). Sweet potatoes were occasionally infected with more than one of these viruses. This is the first report of SwPLV and SPCSV in Korea.

• 디지털융복합연구
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• 제13권8호
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• pp.503-514
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• 2015

은행잎 유래 플라보놀 성분이 갖는 항아토피 활성에 대해 입증된 바는 드물다. 이 논문에서는 MC/9 비만세포에서의 은행잎 플라보놀의 항아토피 효과를 조사하기 위하여 ELISA와 Real-time PCR, western blot으로 은행잎 플라보놀을 분석하였다. 분석 결과 IL-13, MIP-1a의 생성량은 농도 의존적으로 현저하게 감소되었으며 IL-4, IL-5, IL-13, TNF-a 유전자 발현량은 25, 50, $100{\mu}g/m{\ell}$의 농도에서 효과적으로 억제되었다. western blot 분석 결과 c-jun과 NFAT-1단백질 발현이 감소되었음을 확인하였다. 이러한 결과들은 은행잎 유래 플라보놀 성분이 NFAT-1, c-jun 전사인자의 전사를 억제함으로써 MC/9 비만세포에서의 Th2 싸이토카인의 생성을 감소시키는 효과를 갖는 것을 나타낸다. 따라서 은행잎 유래 플라보놀 성분이 아토피 피부염 치료제로서 활용가능성이 있음을 보고하고자 한다.

• 식물병연구
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• 제18권3호
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• pp.231-235
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• 2012

본 연구는 citrus에 심각한 피해를 초래하는 바이러스인 ICRSV가 국내로 유입되는 것을 차단하여 그로 인한 피해를 방지하기 위해 이를 진단하는 시스템을 구축하고자 하였다. ICRSV가 감염된 시료를 구할 수 없어 외피단백질 유전자를 E. coli의 codon usage를 고려하여 optimization한 뒤 E. coli에서 수용성 단백질로 과발현된 재조합 ICRSV 외피단백질을 정제하였다. 정제한 재조합 단백질을 이용해 제작한 복클론 항체는 $1{\times}10^{-4}$으로 희석하였을 때 western blot과 ELISA를 통해서 각각 10 ng, 5 ng의 재조합 ICRSV 외피단백질을 검출할 수 있었다. 이로써 제작된 항체를 이용하여 소량의 바이러스 입자만으로도 ICRSV를 검출할 수 있을 것이다.

• 대한수의학회지
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• 제35권1호
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• pp.143-148
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• 1995

Brucella abortus cell wall antigen was extracted from Brucella abortus 1119-3 by ultrasonication and followed by sodium dodecyl sulfate(SDS) treatment. In order to confirm whether this preparation is serologically cross reactive with Yersinia enterocolitica 0:9, Western blot analysis with mouse anit-Brucella abortus1119-3 and with mouse anti-Yersinia enterocolitica 0:9 was performed. ELISA results from using those Brucella antigen and Yersinia antigen were assessed whether they had correlation. According to the results of western blot analysis and ELISA, there was no evidence of cross reactivity between the Brucella abortus 1119-3 antigen preparation and Yersinia enterocolitica 0:9. Therefore the SDS treated antigen prepared in this study could be suitably used as specific ELISA antigen without confusion in the interpretation of serological tests for brucellosis in cattle.

• Journal of Korean Dental Science
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• 제11권2호
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• pp.62-70
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• 2018

Purpose: Bone marrow has long been a source of primary cells. This study was performed to evaluate the effects of age and sex on the cellular viability and expression of stem cell markers of mRNA and on the protein expression of bone marrow stem cells (BMSCs) derived from healthy donors. Materials and Methods: Stem cells were isolated from human bone marrow and plated in culture plates. The shape of the BMSCs was observed under inverted microscope. Quantitative cellular viability was evaluated using a Cell-Counting Kit-8 assay. The expression of stem cell surface markers was tested and a series of quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot was performed to evaluate the expression in each group. Result: The shapes of the cells at 20s, 30s, and 50s were similar to each other. No significant changes in cellular viability were noted among different age groups or sex groups. The BMSCs expressed CD44, CD73, and CD90 surface markers but did not express CD14 and CD34. There were no noticeable differences in CD surface markers among the different age groups. The expressions of CD surface markers were similar between men and women. No significant differences in the secretion of vascular endothelial growth factors (VEGFs) were noted at Day 3 between different age groups. qRT-PCR regarding the expression showed differences between the age groups. However, Western blot analysis showed a decrease in expression but did not reach statistical significance (P>0.05). Conclusion: This study clearly showed no significant differences in shape, cell viability, expression of stem cell surface markers, or secretion of human VEGF among different age groups. However, western blot analysis showed a tendency of age-related decrease which did not reach statistical significance. Collectively, autologous or allogeneic BMSCs should be meticulously applied to obtain optimal results regarding age and sex.

• Biomolecules & Therapeutics
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• 제8권2호
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• pp.107-112
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• 2000

To investigate the alteration of airway mucin in airway disease patients, immunoassay procedures were employed using monoclonal antibodies HM02 and HM03 (Hybridoma, 18,457-463, 1999). Alteration of mucin release was determined by ELISA and the integrity of mucin was determined by Western blot. In ELISA, it was found that mucin release increased from pneumonia, chronic cough, bronchiectasis, eosinophilic pneumonia, lung cancer and bronchial asthma patients. In Western blot, the increase in immunoreactivity was observed in case of pneumonia, chronic cough, bronchiectasis and bronchial asthma. In bronchial asthma, there was no obvious degradation of mucin while in other diseases, varying degree of mucin degradation was observed. The data from the present study implicate that HMO2 and HM03 are suitable for the immunological analysis of mucin in airway disease patients. The role of increased mucin release and varying degree of mucin degradation on airway diseases should be further investigated in the future.

• 한국균학회지
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• 제39권3호
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• pp.235-238
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• 2011

팽이균사체의 형질전환을 위하여 Agrobacterium 세포를 사용하였다. 특히, Agrobacterium 세포의 감염단계 전에 약한 NaOH용액을 처리하였으며 이로써 균사체 세포들의 표면 상해 발생을 기대하였다. 그 결과, hygromycin 저항성 ($hyg^r$) 균사체는 NaOH 처리를 거친 경우에서만 출현하였다. 형질전환 균사체의 $hyg^r$ 유전자 도입은 PCR로 확인되었으며 또한 Southern blot hybridization과 western blotting 분석에 의하여 단일 유전자 copy의 삽입과 외래유전자의 발현을 확인할 수 있었다. 본 연구는 팽이균사체에 대한 효율적인 Agrobacterium 이용 형질전환수단을 보여주고 있다.

• 동의생리병리학회지
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• 제23권1호
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• pp.199-205
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• 2009

It is unclear whether Fructus ligustri Lucidi (FLL) extract anti-proliferative effect in human glioma cells. The present study was therefore undertaken to examine the effect of FLL on cell viability and to determine the underlying mechanism in A172 human glioma cells. Cell viability and cell death were estimated by MTT assay and trypan blue exclusion assay, respectively. Apoptosis was measured by Annexin-V binding assay and cell cycle analysis. Activation of kinases and caspase-3 was estimated by Western blot analysis. FLL resulted in apoptotic cell death in a dose- and time-dependent manner. FLL-induced cell death was not associated with reactive oxygen species generation. Western blot analysis showed that FLL treatment caused down-regulation of PI3K/Akt pathway, but not ERK. The PI3K/Akt inhibitor LY984002 sensitized the FLL-induced cell death and overexpression of Akt prevented the cell death. FLL induced caspase-3 activation and the FLL-induced cell death was prevented by caspase inhibitors. These findings indicate that FLL results in a caspase-dependent cell death through a P13K/Akt pathway in human glioma cells. These data suggest that FLL may serve as a potential therapeutic agent for malignant human gliomas.

• 한방안이비인후피부과학회지
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• 제22권2호
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• pp.1-18
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• 2009

Objective : Moutan Cortex (the root bark of Paeonia suffruticosa Andr.) is widely used in oriental medicine as a remedy for inflammation. However, as yet there is no clear explanation of how MC(Moutan Cortex) affects the production of inflammatory cytokine. This study was to determine the effects of Essence extracted MC on the mast cell-mediated inflammatory responses. Method : We observed the effect of MC on compound 48/80-induced histamine release of rat peritoneal mast cells and the effect of administering MC on PCA in rat. We measured the amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A23187) in the human mast cell line (HMC-1) incubated with various concentrations of MC. The TNF-$\alpha$ protein levels were analysised by Western blot. The TNF-$\alpha$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The TNF-$\alpha$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. NF-$\kappa$B, phospho-I$\kappa$B and MAPKs were exmined by Western blot analysis. The NF-$\kappa$B promoter activity was examined by luciferase assay. Result : 1. Enzyme immunoassay indicated that MC suppressed histamine secretion of rat peritoneal mast cells. 2. In PCA dependent on IgE, MC had anti-allergic effect of the internal surface of rat skin. 3. Western blot indicated that MC decreased TNF-$\alpha$ protein levels. 4. ELISA indicated that MC decreased TNF-$\alpha$, IL-6 but MC had no significant effect on IL-8 in HMC-1 cells. 5. RT-PCR indicated that MC decreased TNF-$\alpha$, IL-8 but MC had no significant effect on IL-6 in HMC-l cells. 6. Western blot indicated that MC suppressed the induction of MAPKs, NF-$\kappa$B & phospho-I$\kappa$B activity in HMC-1 cells. 7. Luciferase assay indicated that MC suppressed the PMA plus A23187-induced NF-$\kappa$B promoting activityin HMC-1 cells. Conclusion : In this study, we have found that MC is an inhibitor of NF-$\kappa$B, MAPKs & cytokines on the mast cell-mediated inflammatory responses.