• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.261-261
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• 1994

Bovine milk xanthine oxidase (E.C.1.1.3.22, XO) purchased from Sigma Chemical Co. had the three protein fragments below 150 kDa on 7.5％ SDS-PAGE, which did not show enzyme activity. To remove these fragments, the enzyme preparation was further purified through Sephadex G-200 column chromatography. Two peaks exhibiting enzymatic activity were separated very closely to the void volume, which were revealed as two different enzyme forms, dimeric and monomeric, confirmed by activity staining on native PAGE. Anti sera-against each of the two enzyme forms were raised by subcutaneous injection at multiple sites on the back of rabbits during 4 weeks. On the immunodiffusion test, it was found that both of the antisera of the two forms could react with each other, which implied that their epitopes were identical In the Western blot analysis of mouse liver cytosol fraction, it was found that rabbit anti-XO antibody bound well with the protein band of monomeric mouse liver XO of about 150kDa. Based on this result, mouse liver cDNA 1 ibrary was screened by in situ hybridizat ion wi th rabbi t anti -XO antibody as probe. Through the immunological screening, recombinant phages giving positive signal by the production of XO were selected and further purified. To validate these clones, purified phages were lysogenized in E. coli Y1089 and their lysates were analysed for enzyme activity and immunoreactivity, It was verified that lysates of the purified recombinant phage lysogens exhibited the enzymatic activity as well as bound wi th XO antibody, when induced by IPTG. The above results assert that selected recombinant phage carries mouse liver XO gene.

• Journal of Sericultural and Entomological Science
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• v.30 no.2
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• pp.96-104
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• 1988

Vitellin, the major yolk protein of the silkworm, Bombyx mori was pruified, and its immunological properties and the quantitative changes during embryogenesis were studied. The ovary transplantation into male hosts was also carried out to find its effect on the yolk protein synthesis. The pupal vitellogenin and the egg vitellin of Bombyx mori were purified by DEAE-cellulose column chromatography. These two female specific proteins showed the same mobility in the polyacrylamide gel electrophoresis and the same reaction in the double immunodiffusion test. The immunological identity was also observed between the vitellins of Bombyx mori and Bombyx mandarina. The rudimentary ovaries transplanted into the male hosts of silkworms produced eggs without vitellin, indicating that the yolk precursors synthesized in other female organ beyond the ovary were necessary to produce vitellins. The major yolk protein, vitellin was disintegrated and utilized mostly during late stage of embryogenesis. It was different characteristics from the egg specific protein, which was utilized continuously from the early embryonic stage.

• Korean Journal of Poultry Science
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• v.26 no.3
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• pp.179-188
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• 1999

The present was undertaken to establish a model for the control of adipocytes differentiation by using antibody from egg yolk. The emulsion of membrane protein of 3T3L-1 cell membrane protein with the complete Freund's adjuvant was firstly immunized in layer. Second and third boosting were undertaken with two weeks intervals by injection of the emulsion of the same antigen with the incomplete Freund's adjuvant. After 4 week of the first immunization, eggs were collected and antibody (IgY) was purified from egg yolk. The purity of IgY was 60-98% determined by single radial immunodiffusion (SRID) methods. Titer value of the antibody showed high reactiviy for the preadipocytes membrane protein measured by ELISA. When the IgY was added in the test media containing either 2.5% porcine serum or 10% FBS(control), the differentiation of 3T3L-1 cells and Glycerol-3-phosphate dehydrogenase(GPDH) activities was significantly decreased compared to the control cells(p〈0.05). When mice were subcutaneously injected with IgY raised against membrane protein of 3T3L-1 cells for 3 weeks, adipose tissue mass around ovary was tended to be decreased in female mice compared to those of control mice. It is suggested that a potential for manipulating of lipid accumulation through decrease in 3T3L-1 cell differentiation and fat accumulation in female mice by IgY treatment.

• Journal of Food Hygiene and Safety
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• v.3 no.2
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• pp.75-81
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• 1988

Various methods such as lel-mtration on Sephadex G-SO, 75, 100 Sephacry, and Ultro gel, and lon-exchanle chromatoaraphy on Amberilte and carboxymethyl (CM)-cellulose, and Fast Protein liquid Chromatolraphy (FPLC) were applied for the purification of enterotoxin B from Staphylococcus aureus ATCC 14458 and compared one another. lon-exchanle chromatography on Amberllte resin was good enough to remove non-entrotoxln materials in culture, convlnient to use and fast although tbe purity was less tban 70%. However, CM-cellulose showed to be better purity and yield tban those of Amberilte resin. The yields of these two resins for ion-exchange cbromatograpby were about 70% and 75%, respectively. When the gel-filtration methods on Sepbadex G-50, 75, 100, Sepbacryl, and Ultro lei were applied, the purities were about 90%. FPLC was found to be tbe most efficient metbod in terms of purity (96%) and speed. For the purification of sample with large volume, particularly, tbe combined metbod, gel-mtration after Amberlite can be also used efficiently. Tbe purified toxin was found to be identical to type B enterotoxin used for reference standard by Oucbterlony immunodiffusion test.

• The Journal of the Korean Society for Microbiology
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• v.18 no.1
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• pp.99-110
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• 1983

Four cases of pulmonary pseudallescheriasis in patients with healed pulmonary tuberculosis are described. All four patients had a long history of antituberculous chemotherapy for pulmonary tuberculosis, but continuous sputum negativity for acid fast bacilli indicated apparent recovery from tuberculosis. They, however, complained continued intermittent hemoptysis and chronic cough. Although their chest roentgenograms did not show a clearcut mycetomal shadows in preformed cavitary lung lesions, Pseudallescheria boydii or Scedosporium apiospermum was repeatedly isolated from serial sputum specimens collected at different days for a period of over half an year or two years and their serial serum specimens produced precipitin bands with home-made antigen from 8-week old culture filtrate of P. boydii. Second fungus was isolated from sputum specimens of two patients and one was Candida albicans and the other was Aspergillus fumigatus. Sera from both patients reacted with antigens of those second fungi. Unfortunately pulmonary function of three patients did not allow surgical excision of the infected area and one patient refused surgery.

• The Journal of the Korean Society for Microbiology
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• v.18 no.1
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• pp.23-29
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• 1983

An epidemiological survey was carried out to identify the serotypes of Streplococcus mutans isolated from carious lesions of 65 caries-active subjects(CAS) and sound tooth surfaces of 40 caries-free subjects(CFS). The autoclaved antigen extract was performed on each isolate, and then, serotypes of the isolates were determined in agar-gel immunodiffusion test. The results were as follows: 1. S. mutans was found in 78% of the samples of CAS, and of CFS. The difference of isolation frequency between CAS and CFS was not observed. 2. Only one serotype per single subject was detected in 61% of total samples, in remaining 39% of samples two or more serotypes were detected. 3. In 41.2% of CAS samples plural serotypes of S. mutans were found, whereas 35.5% of CFS samples showed plural serotypes distribution. 4. The most frequently identified serotype in each subject was serotype c; 69.5% of subjects harbored serotype c S. mutans. Serotype d was next most frequently isolated from subjects, comprising 23.2%. 5. Serotype c strain was found in 64.7% of CAS, 77.4% of CFS. 6. Of the isolates from CAS and CFS, serotype c was most commonly found, comprising 48.8%, serotype d was found in 16.3%, serotypes f. e, and g comprising 13.2%, 9.3%, and 7.8% respectively. Serotypes a and b were also found but in far lower frequencies(2.3%, 0.8%). 7. Serotype c strains were more found in CFS than in CAS, but serotypes d and e were more found in CAS.

• Restorative Dentistry and Endodontics
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• v.8 no.1
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• pp.31-44
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• 1982

Streptococcus mutans strains were isolated from dental plaques of carious lesions of 53 patients on mitis-salivarius-bacitracin (MSB) and mitis-salivarius(MS) medium as a supplement. The epidemiological investigation was carried out to determine the biotypes and serotypes of S. mutans isolates. For the serotyping, autoclaved extract antigens from the isolates and serotype-specific antisera against seven known serotypes of S. mutans were prepared. The serotypes of the isolates were demonstrated in immunodiffusion test. In addtition, the prevalence of ${\beta}$-hemolysis on 5% sheep blood agar plate in restricted anaerobic condition and yellow pigment production on 5% sucrose agar plate in less anaerobic condition among the isolates were investigated. The results were as follows: 1. Forty-eight S. mutans strains were isolated from dental plaque samples of 33 patients (62.3%) among 53 patients. 2. Of the isolates, some strains were not grown on MSB medium. 3. Serotype c S. mutans was found in 60.6%, serotype d was found in 30.3% of the patients who were known to harbor S. mutans. 4. Of. the isolates, serotype c isolates were most prevalent (43.8%), serotype d isolates were 25.0%, and serotype b, e, f and g isolates were also found but in lower frequencies. Serotype a S. mutans were not detected. 5. The correlation between serotype and biotype of the isolates was found in 78.6% of the typing cases. 6. Strains revealed ${\beta}$-hemolysis were in 52.1% of the isolates, strains produced yellow pigment were in 47.9% of the isolates, and with one exception, all the strains were belong to serotype c, e and f. 7. The majority of the isolates which revealed ${\beta}$-hemolysis appeared to be yellow pigmented, these isolates were belong to serotype c, e and f.

• Food Science of Animal Resources
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• v.25 no.1
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• pp.98-102
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• 2005

The purpose of this study was to demonstrate biochemical properties of lactoferrin (Lf) obtained from the colostrum of Korea native cow. The molecular weight of the purified Korean native cow's Lf (K-Lf) was 81kDa, the isoelectric point was 9, and the content of iron was 0.56 mg/g, which is indicated that iron saturation of the lactoferrin was 40.6%. Amino acid composition and a-helix content were different K-Lf from bovine Lf (B-Lf). Immunological cross reactivity was observed between K-Lf and B-Lf but not between K-Lf and human Lf (H-Lf) by immunodiffusion test and Western blot analysis. Out results indicate that structure of K-Lf is different from that of B-Lf although K-Lf and B-Lf were immunologically cross-reactive.

• Korean journal of applied entomology
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• v.22 no.4 s.57
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• pp.293-299
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• 1983

Experiments were carried out to elucidate the specific interactions between host plant, Phaseolus vulgaris, and symbiotic bacteria, Rhizobium Phaseoli. Purified P. vulgaris lectins and six species of cultured Rhizobium were subjected to agglutination test. Lectins from bean and R. phaseoli showed relatively high agglutination activity indicating that host plant lectins recognize carbohydrate moieties on the compatible Rhizobium cell surface. The specific carbohydrate receptors for binding of the lectins on the cell surface of R. phaseoli were found as mannose and galactose. The minimum concentration of sugars for the inhibition was 6.25mM. The lectin content of cultured plant roots was measured after germination and was maximum in 5-day seedlings. The nodulation was competitively inhibited by lectins for the plants cultured with Rhizobium cells. By immunochemical studies, there was some relationship in antigenic determinants between R. phaseoli and R. japonicum but no relationships were observed with other Rhizobium species. The results suggest that the infection by rhizobia to the roots of leguminous plants may be caused by the specific interaction of lectins with rhizobia.

• Applied Biological Chemistry
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• v.41 no.7
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• pp.522-526
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• 1998

Ferritin from germinated soybean seeds was purified by ammonium sulfate precipitation (0.55 saturation), ion-exchange chromatography on DEAE-cellulose, gel filtration chromatography on Sephacryl S-300, and HPLC with Bio-Scale Q2 column. SDS-PAGE analysis showed that the purified ferritin is composed of subunit with an apparent M, 21,000. The molecular mass of the native soybean ferritin estimated by gel filtration on Sephacryl S-300 and non-denaturing polyacrylamide gel electrophoresis appeared to be $510{\sim}560\;kDa$. Soybean ferritin contained 833 mol Fe/mol protein, which is 31-fold more iron than pumpkin ferritin and stained positive for iron on non-denaturing gel. Soybean ferritin cross-reacted with anti-soybean rabbit ferritin antiserum.