• 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.

• Journal of Microbiology and Biotechnology
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• 제2권4호
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• pp.237-242
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• 1992

In order to clone the gene coding for alkaline phosphatase in the yeast Kluyveromyces fragilis, a genomic library was constructed using the yeast-E. coli shuttle vector pHN114 as a cloning vector. From the genomic library, a clone carrying the gene was isolated and the plasmid was designated as pSKH101. A restriction enzyme map was made using this plasmid. Subcloning experiments and complementation studies showed that alkaline phosphatase was active only in the original 3.1 kb insert. Southern hybridization analysis confirmed that the cloned DNA fragment was derived from K. fragilis genomic DNA. Using a minicell experiment, the product of the cloned gene was identified as a protein with a molecular weight of 63 KDa. A 0.6 kb HindIII fragment, which showed promoter activity, was isolated using the E. coli promoter-probe vector pKO-1.

• 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.

• Nutritional Sciences
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• 제7권1호
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• pp.47-52
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• 2004

The food composition database is used to evaluate the nutritional value of foods in Korea. It is used as a source for basic data in many fields, not only for the establishment of the National Food Supply Plan but also for nutritional research, to aid in the drafting of the National Food Policy, and in clinical and epidemiological research. The database is also used by institutional and commercial food services and by the food processing industry among others. In 1970, the Rural Development Administration(RDA) published the "Korean Food Composition Table" for the first time and it has since become the established authority on food composition for the entire nation. The latest published food composition table (the 6th edition, published by the National Rural Living Science Institute, KDA) covers 2,337 kinds of foods and consists of two volumes. The National Rural Living Science Institute, the Korea Food and Drug Administration, and the National Fisheries Research and Development Institute have continuously improved production of food composition data in Korea. As consumers have grown more interested in the quality and safety of foods and raw agricultural products, we can expect even further development of food composition data through improvements in quality and increases in quantity. We need to develop more comprehensive food composition data by diversifying the kinds of foods that are included, unifying analytical methods, and increasing the number of nutrients covered. Moreover, food composition data must be modified in other ways to make for a better food composition database for the convenience of users. of users.

• 생명과학회지
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• 제8권5호
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• pp.598-603
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• 1998

1. 어류에서 분리한 V. anguillarum KS410의 용혈소를 분리하여 배양 상등액, 황산암모늄 침전, DEAE-cellulose column chromatography, Sephadex G-200 gel 여과 등의 과정을 통하여 36배 정제하였고 최종 회수율은 2.3%였다. 2. 정제된 용혈소의 SDS-PAGE상의 전기영동 결과는 38 Kda의 분자량을 보유하였다. 3. 온도에 대한 정제 용혈소는 45$^{\circ}C$까지는 안정하였으나 그 이상의 온도에서는 급격히 실활되어 이열성인 것으로 나타났다. 4. NaCl 농도에 따른 V. anguillarum 용혈소의 안정성은 1%가 가장 안정하였고 4%이상에서는 안정성이 감소하였다. 5. 정제된 용혈소의 pH 안정성은 6-9 사이었다. 6. 금속이온에 대한 정제 용혈소는 $Ca^{2+}, Cu^{2+}, Zn^{2+}, Fe^{2+}$ 첨가시에는 실활되었으나 $Mg^{2+}$첨가에는 영향을 받지 않았다. 7. 정제용혈소는 EDTA에 대하여 전혀 활성에 저해를 받지 않았으나 $FeCl_3$에서는 저해를 받았다.

• 한국식물병리학회지
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• 제14권1호
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• pp.62-67
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• 1998

Barley yellow mosaic virus (BaYMV-HN) occurring Haenam area was isolated by mechanical inoculation onto barley cultivars, purification and production of antibody. BaYV-HN were purified from infected plants a filamentous virus with 13 nm in diameter and 250∼300 nm and 500∼650 nm in length. Specific antibody made by injecting the purified virus to the muscle of a rabbit. In gel-diffusion tests antibody to BaYMV-HN did not make spur with tow Japanese BaYMV isolates BaYMV-II-1 or BaYMV-III. BaYMV-HN showed the symptom of yellowing and necrosis in host plants. Mechanical inoculation tests with Japanese barley cultivars showed that BaYMV-HN infected New Golden, Akagi Nijo and Tosan Kawa 73, but did not infect Amagi Nijo, Haruna Nijo, Ishukushirazu (ym3), Misato Golden (Ym1), Kashimamugi, Joshushiro Hadaka and Mokusekko 3 (ym1). In Korean barley cultivars, some of the naked barleys which are Hinssalbori, Kinssalbori, Saessalbori and Saechalssalbori were not infected by BaYMV-HN. However, it infected all the covered barley cultivars and the beer barley cultivars. BaYMV-HN had two RNAs, RNA 1 (7.6 Kb) and RNA 2 (3.5 Kb), and one coat protein (33 KDa).

• 한국식물병리학회지
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• 제14권1호
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• pp.68-73
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• 1998

Barley mild mosaic virus (BaMMV-Kor) was isolated from the southern part of Korea, and by mechanical inoculation onto barley cultivars, purification and production of antibody. BaMMV-Kor purified form infected plants were filamentous particle, with 13 nm in diameter and 250∼300 nm and 500∼650 nm in length. Antibody of BaMMV-Kor was made by injecting the purified virus to the muscle of a rabbit. In gel-diffusion test, antibody to BaMMV-Kor created spur with BaMMV-Kal and BaMMV-M, but did not make spur with BaMMV-Kor infected New Golden, Ishukushirazu, Joshushiro Hadaka and Misato Golden, but did not infect Kashimamugi, Chikurin Ibaraki 1 and Mokusekko 3. In Korean barley cultivars, BaMMV-Kor infected most of the covered barley cultivars, but did not infect Saeolbori. It also infected naked barley cultivars except Chalbori and Hinssalbori. And all the beer barley cultivars were infected by BaMMV-Kor. BaMMV-Kor had two RNAs, RNA 1 (7.5 Kb) and RNA 2 (3.5 Kb), and coat protein (33 KDa).

• 한국식품조리과학회지
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• 제20권4호
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• pp.387-395
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• 2004

The effect of commercial Doenjangs (Korean soybean pastes) on sponge cake making was studied. When Doenjang was added to e99 foams at levels of 5, 10, 15 and 20%, there were no big differences in viscosity or stability of egg foams up to 15%, and thus in cake volume index, compared to those of control (without Doenjang). In the results of preference rank test, cake with 10% Doenjang was preferred to the other cakes. Therefore, it was decided that the addition of 10% Doenjang was suitable for sponge cake making. When seven commercial Doenjang products were used at 10%, there were differences in physical and sensory properties. The decreased viscosity of egg foams reduced their stability and so decreased the quality of the sponge cakes with low volume index and low texture scores. The use of Doenjangs improved the flavor of sponge cake and so countervailed the undesirable egg flavor. Especially, “C” Doenjang product, which increased the viscosity and stability of egg foams, produced good foaming ability and the highest physical and sensory quality, thus the higher overall acceptable scores than the control. It was probably due to its high amount of protein hydrolysate above 10-KDa. Consequently, the suitable amount of Doenjang was fixed at 10% and “C” Doenjang product was chosen as the best among the commercial products for sponge cake making.

• 한국작물학회지
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• 제44권1호
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• pp.78-85
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• 1999

To investigate kernel hardness, a compression test which is widely used to measure the hardness of individual kernels as a physical testing method was made simultaneously with the measurement of friabilin (15KDa) which is strongly associated with kernel hardness and was recently developed as a biochemical marker for evaluating kernel hardness in 79 Korean wheat varieties and experimental lines. With the scattered diagram based on the principal component analysis from the parameters of the compression test, 79 Korean wheat varieties were classified into three groups based on the principal component analysis. Since conventional methods required large amount of flour samples for analysis of friabilin due to the relatively small amount of friabilin in wheat kernels, those methods had limitations for quality prediction in wheat breeding programs. An extraction of friabilin from the starch of a single kernel through cesium chloride gradient centrifugation was successful in this experiment. Among 79 Korean wheat varieties and experimental lines 50 lines (63.3%) exhibited a friabilin band and 29 lines (36.7%) did not show a friabilin band. In this study, lines that contained high maximum force and the lower ratio of minimum force to maximum force showed the absence of the friabilin band. Identification of friabilin, which is the product of a major gene, could be applied in the screening procedures of kernel hardness. The single kernel analysis system for friabilin was found to be an easy, simple and effective screening method for early generation materials in a wheat breeding program for quality improvement.

• 대한수의학회지
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• 제36권3호
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• pp.615-619
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• 1996

Sera of pigs with clinically normal and infectious conditions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) to demonstrate the specific changes in protein profile. In the sera from pigs with infection, haptoglobin with a 40KDa protein was found to be increased as compared to that of sera from normal pigs. As a rapid detection method for monitoring infections at large-scale farms, one of acute phase protein, haptoglobin, was selected to compare the concentrations between infectious and non-infectious conditions. Haptoglobin concentrations were low in pigs with clinically normal conditions but significantly increased in pigs with Aujesky's disease, hog cholera and parvo-virus infection. The studies provide that haptoglobin can be used as an indicator to monitor infections early at farm level.