• 한국축산식품학회지
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• 제28권4호
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• pp.499-504
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• 2008

Microwave,초음파 및 초고압 처리에 의한 PSA의 항원성 변화를 살펴본 결과는 다음과 같다. microwave처리에 의한 변화를 알아본 결과 처리시간이 증가함에 따라 PSA와 항체와의 결합력이 감소하였으며 특히 10분 처리 시에 약 30% 정도로 가장 낮은 결합력을 나타내었다. SDS-PAGE 결과에서는 5, 10분 처리 시에 PSA band가 많이 약화되었으며 immunoblotting에서도 5, 10분 처리한 PSA가 항체와 거의 반응하지 않았다. 하지만 열을 배제하고 microwave 처리한 경우에는 처리구 모두 큰 변화 없이 높은 결합력을 유지하였다. 초음파 처리(5, 10, 30 및 60분)에 의한 PSA의 항원성 변화를 살펴본 결과에서는 초음파 처리구 모두 약 78% 정도의 높은 결합력을 보였으나 immunoblotting에서는 초음파 처리한 PSA가 항체와 거의 반응하지 않았다. PSA에 초고압 처리(200, 300 및 400 MPa)한 후 항원성 변화를 살펴본 결과 초고압 처리구 모두 PSA와 항체와의 결합력이 약 98%로 높은 결합력을 유지하였으며 immunoblotting에서도 초고압 처리구의 PSA가 항체와 강하게 결합하였다. 이상의 결과를 종합해 볼 때 열을 배제한 microwave, 초음파 및 초고압 처리는 PSA의 항원성을 감소시키지 못하였으나 열을 배제하지 않은 microwave 처리는 시간이 증가함에 따라 PSA의 항원성을 크게 감소시켰으므로 이를 저 알레르기 돼지고기 식품 개발에 응용할 수 있을 것으로 사료된다.

• Biomolecules & Therapeutics
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• 제9권1호
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• pp.7-14
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• 2001

The chicken meat has been reported as one of the food causing allergic reactions predominantly to Korean. At present, several in vitro tests for immunoglobulinG (IgG)-mediated as well as IgE-mediated food allergy are available. 13 clinically chicken meat-allergic patients were investigated together with 4control subjects for identification of chicken meat-specific reactivity by ELISA. Also, protein profile and IgE, IgGtotal and IgG4-reacting allergens were detected by means of sodium dodecyl sulfate-polyacrylamide gel electro-phoresis (SDS-PAGE)and immunoblotting. Chicken meat extracts were prepared as raw, heated, heat and simulated gastric fluid (SGF) treated samples to characterize the stability of allergen to physicochemical treatment. SDS-PAGE revealed 9~200 kDa bands. And in immunoblotting 7 sera were identified most major bands between 10 and 78 kDa. In case of IgE, six proteins (17, 26, 35, 40, 78 kDa) were predominant in heat-treated extract, and the one (35 kDa) was present in SGF-treated preparations. In case of IgG$_{total}$ and IgG4, most of them showed a patters simmilar to IgE. There were significant differences (P<0.05) in IgE, IgG$_{total}$ , IgG4 Abs to chicken meat between the allergic and control subjects in ELISA. In addition, the concentration of IgG4Abs in the challenge-positive subjects was significantly higher than that of control subjects. It is considered that the specific IgE response to chicken meat was rarely prevalent to Koreans. However, the specific IgG4 response play an important role in the development of allergic symptoms.

• 대한의생명과학회지
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• 제8권3호
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• pp.107-113
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• 2002

Enterohemorrhagic Escherichia coli (EHEC) strains of serotype O157:H7 have been shown to colonize the intestinal epithelial cell by the attaching and effacing (AE) mechanism. The AE lesion is mediated by an intimin, of which production and expression are controlled by a 3-Kb eaeA gene located EHEC chromosomal DNA. If the eaeA gene is mutated, EHEC O157:H7 strains lose capacity of adhesion to intestinal epithelial cells. In this study, a 891 bp of the 3'-end region of a gamma intimin was amplified by polymerase chain reaction (PCR). The PCR product was inserted into pSTBlue-1 cloning vector and transformed into DE3 (BL21) competent cell. After plasmid mini-preparation and restriction enzyme digestion of eaeA/891-pSTBlue-1 vector, target eaeA gene was re-inserted into pET-28a expression vector and was transformed. Then the expression of recombinant eaeA/891 (891 bp) gene was induced by isopropyl-$\beta$-D-thiogalactopyranoside (IPTG). The expression of the 40-KDa recombinant protein was identified in SDS-PAGE and confirmed by immunoblotting using the His.Tag$^{\circledR}$ and T$_{7}$.Tag$^{\circledR}$ monoclonal antibody. This recombinant protein expressed by eaeA gene could be applied in further studies on the mechanisms of E. coli O157:H7 infection and the development of recombinant vaccine.

• The Korean Journal of Physiology
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• 제25권2호
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• pp.115-123
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• 1991

Molecular association of glucose transporters with the other proteins in the plasma membrane was assessed by gel electrophoresis and immunoblot techniques. Approximately $31.5{\pm}5.1%$ of GLUT-4, $64.8{\pm}2.7%$ of clathrin, 48.7% of total protein in the plasma membrane (PM) were found insoluble upon extraction with 1% Tx-100. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the Tx-100 insoluble PM fraction contained about 4 major polypeptides with apparent molecular weight of above 200, 100-120, 80 and 30-35 KDa that were readily removed upon wash with a high pH buffer which is known to remove clathrin and 0.5 M Tris-buffer which is known to remove assembly proteins (AP). Immunoblotting of GLUT4 and clathrin against specific antibodies showed that GLUT-4 and clathrin were co-solubilized up to 84.6% and 82.7% respectively by wash with a high pH buffer and 1% Tx-100. When the membrane was pre-washed with a high pH buffer and 0.5 M Tris solution, GLUT4 and clathrin were not solubilized further suggesting that GLUT4 molecules are in molecular association with clathrin, AP and/or other extrinsic membrane proteins in plasma membrane and the formation of clathrin-coated structures might be involved in insulin stimulated glucose transporter translocation mechanism.

• Parasites, Hosts and Diseases
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• 제46권1호
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• pp.29-32
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• 2008

The aim of this study is to determine the Toxocara seropositive rate among healthy people with eosinophilia. A total of 97 people residing in Seoul who were healthy and whose blood eosinophilia was over 10%, as shown by regular health check-ups in 2004, were subjected to this study. Their sera were tested by immunoblotting and ELISA with the antigen of larval Toxocara canis excretory-secretory (ES) protein. Sixty-five sera were band-positive (67.0%). The seropositve control sera were positive to band sizes of 66 kDa, 56 kDa, 32 kDa, and 13 kDa. In ELISA, 63 sera (65.0%) were positive to T. canis ES protein. There was no significant correlation between the IgG ELISA titer and the level of eosinophilia (r = 0.156, P = 0.156). As there were insufficient data to determine whether there were cross-reactions with other helminthic infections, or whether atopy occurred, further studies are required to verify the cause of the seropositive reactions against T. canis ES antigen. Toxocariasis seropositivity is suggested to be the major cause of eosinophilia, since the Toxocara seroprevalence among Korean rural adults was shown to be approximately 5%.

• Natural Product Sciences
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• 제6권3호
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• pp.147-150
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• 2000

The extracts of Carica papaya (flower), Barringtonia macrostachya (leaves), Coleus tuberosus (tuber), Mangifera indica (fruit skin) and Eugenia polyantha (leaves) showed strong in vitro anti-tumor promoting activity when assayed using Raji cells (Mooi et al., 1999). The antitumor promoting activity of the crude extracts was further analyzed by immunoblotting analysis of Raji cells carving Epstein-Barr virus genome. The expression of early antigens diffuse (EA-D) and early antigens restricted (EA-R) was determined by performing western blotting of treated Raji cells with human sera of nasopharyngeal carcinoma patients. All the plant extracts were shown to be able to suppress both EA-D and EA-R.

• 한국가축번식학회지
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• 제14권2호
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• pp.125-131
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• 1990

These experiments were carried out to investigate of the molecular characteristics of porcine zona pellucidae and to examine the reactivity of anti-zona antibody by SDS-PAGE, Immunoblotting and Immunoprecipitation. The results obtained in these experiments were summarized as follows : 1. The result obtained by SDS-PAGE of porcine zona pellucidae indicated that it composed of several units with molecular weight ranging 55,000-110,000. 2. In order to see the reactivity of antibodies to zona pellucidas, immunoblotting was applied. The results indicated that polyclonal antibodies to porcine and mouse zona reacted with porcine zona. While monoclonal antibody to porcine did not react with the procine zona enough to show a clear band on a gel. 3. Labelling of porcine zonae with 125I was performed using the Iodogen method, the radioactivity and the percent incorporation of 125I into porcine zonae were approximately 26,000 cpm/10${mu}ell$ and 16, respectively. 4. Measurements of radioactivity and O.D value for Immunoprecipitates produced by reaction of 125I-porcine zona with anti-zona antisera were confirmed that existence of reactivity of monoclonal antibody to porcine zona although its reactivity was lower than that of polyclonal antibodies, and reconfirmed that cross-reactivity of polyclonal antibody of mouse zona with porcine zona.

• The Korean Journal of Physiology and Pharmacology
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• 제12권6호
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• pp.331-336
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• 2008

The present study was aimed to determine whether there is an altered regulation of tubular transporters in gentamicin-induced nephropathy. Sprague-Dawley male rats ($200{\sim}250\;g$) were subcutaneously injected with gentamicin (100 mg/kg per day) for 7 days, and the expression of tubular transporters was determined by immunoblotting and immunohistochemistry. The mRNA and protein expression of OAT was also determined. Gentamicin-treated rats exhibited significantly decreased creatinine clearance along with increased plasma creatinine levels. Accordingly, the fractional excretion of sodium increased. Urine volume was increased, while urine osmolality and free water reabsorption were decreased. Immunoblotting and immunohistochemistry revealed decreased expression of $Na^+/K^+$-ATPase, NHE3, NBC1, and AQP1 in the kidney of gentamicin-treated rats. The expression of OAT1 and OAT3 was also decreased. Gentamicin-induced nephropathy may at least in part be causally related with a decreased expression of $Na^+/K^+$-ATPase, NHE3, NBC1, AQP1 and OAT.

• 대한미생물학회지
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• 제35권2호
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• pp.141-147
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• 2000

We purified enolase from Candida albicans KNIH10 strain which was isolated from a clinical specimen in Korea. The purified enolase was used to detect anti-Candida antibodies in sera of patients with invasive candidiasis. For purification of enolase from the crude extract prepared by French pressure at 20,000 PSI, the fast performance liquid chromatography (FPLC) using DEAE-sepharose column was used. The elutes at $0.3{\sim}0.4\;M$ NaCl in FPLC was purified with homogenity in SDS-PAGE and its enzymatic activity was confirmed in sera of invasive candidiasis with candidemia patient by immunoblotting. The purified enolase indicated no signal (100% specificity) in 40 normal human sera and 75% (6/8) sensitivity in sera of candidemic patients with suspicious invasive candidiasis by immunoblotting.

• Food Science and Biotechnology
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• 제18권2호
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• pp.317-322
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• 2009

The proteins from 15 types of cultivar of hot peppers cultivated in Korea were extracted and its allergenicity was investigated by immunoblotting and enzyme-linked immunosorbent assay (ELISA). The immunoblotting of hot pepper proteins extracts (HPEs) against serum of hot pepper sensitized patients revealed dominant IgE binding to 14, 37, and 40 kDa molecules. The specific levels of IgE to HPEs sample No. 1, 3, and 7 were much higher than the other samples in patients. Also, IgE binding capacity of HPEs were not reduced by thermal processing and digestion in ELISA using human IgE antibody acquired from hot pepper sensitized patients. By means of Western blotting using anti-thaumatin IgY, thaumatin-like protein (TLP) acting as allergen in several plants and fruits was detected in tested hot peppers. This study demonstrates that the antigenic protein in hot peppers are present but are differently contained according to cultivars.