• Microbiology and Biotechnology Letters
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• v.20 no.1
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• pp.53-60
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• 1992

Nonregulatory trehalase has been purified from mycelia of Rhztoctonia solani. a pathogen of rice sheath blight. Purification procedures involved sonification, gel filtration and fast protein liquid chromatography. Purity was confirmed by isoelectric focusing with silver staining. The purified trehalase was estimated to have a molecular weight of 54,000 and pI point of 5.1. Maximal activity was observed at pH 5.4 and temperature $45^{\circ}C$. The purified trehalase exhibited on apparent Km for trehalose of 3.11 mM and a Vmax of 105.3 $\mu mol min^{-1}\times mg^{-1}$. Validamycin, a commercial antibiotics of rice sheath blight, was a non-competitive inhibitor of Rhzzoctoniu solani trehalase.

• Journal of Microbiology and Biotechnology
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• v.27 no.8
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• pp.1519-1528
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• 2017

Foodborne contamination and salmonellosis caused by Salmonella Enteritidis (S. Enteritidis) are a significant threat to human health and poultry enterprises. Outer membrane vesicles (OMVs), which are naturally secreted by gram-negative bacteria, could be a good vaccine option because they have many biologically active substances, including lipopolysaccharides (LPS), outer membrane proteins (OMPs), and phospholipids, as well as periplasmic components. In the present study, we purified OMVs derived from S. Enteritidis and analyzed their characteristics through silver staining and sodium dodecyl sulfate polyacrylamide gel electrophoresis. In total, 108 proteins were identified in S. Enteritidis OMVs through liquid chromatography tandem mass spectrometry analysis, and OMPs, periplasmic proteins, and extracellular proteins (49.9% of total proteins) were found to be enriched in the OMVs compared with bacterial cells. Furthermore, native OMVs used in immunizations by either the intranasal route or the intraperitoneal route could elicit significant humoral and mucosal immune responses and provide strong protective efficiency against a lethal dose (~100-fold $LD_{50}$) of the wild-type S. Enteritidis infection. These results indicated that S. Enteritidis OMVs might be an ideal vaccine strategy for preventing S. Enteritidis diseases.

• Fisheries and Aquatic Sciences
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• v.3 no.1
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• pp.52-63
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• 2000

Non-specific reaction has been a problem in doing, especially, research and diagnosis for infectious agents. Avidin-biotin-peroxidase complex (ABC) techniques has widely been used to amplify a reaction. Photobacterium damse1a subsp. piscicdia (formerly Pasteurella piscicida) exhibited a capacity to bind with streptavidin non-specifically. The band, estimated 26 K Da in Western blotted paper, was blocked with biotin but incompletely. In an attempt to explore an involvement of the non-specific substance in attaching piscine cells, cell attachment test performed using anti- Ph. d. subsp piscicida sera raised mouse and rabbit exhibited slightly blocking effects for Mediterranean (1736) and significantly for Japanese (Sp 92144) isolate. Biotin decreased the attachment ability significantly for Sp92144 but it was not effective to 1736. Both isolates showed greatly enhanced attachment ability with poly-L-lysin. The non-specific binding substance was contained in bacterial extracellular products (ECPs). The substance was able to purified with 2-imminobiotin affinity column, the purified substance appeared to have 4 bands in silver staining, and had a carbohydrate branch. This purified substance showed cytotoxic effects selectively between 5 piscine cell lines. Moreover, it stimulated rainbow trout macrophage in terms of reduction of cytochrome cas well as yeast phagocytosis, significantly.

• Korean journal of applied entomology
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• v.30 no.2
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• pp.144-149
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• 1991

Biochemical characteristics of Spodoptera uigua nuclear polyhedrosis virus (SeNPV) isolated in Jinju were studied. SeNPV contained a number of nucleocapsids within a viral envelope embeded in polyhedra. The polyhedral protein of SeNPV was composed of a single polypeptide with a M. W. of 30kd. Double-immunodiffusion test showed that the polyhedral protein of SeNPV had common antigenic determinants with SINPV and BmNPV. Virion proteins of SeNPV were resolved into 49 polypeptides by silver staining after SDS-PAGE. The approximate genome size of SeNPV by restriction endonuclease analysis was 1l0kb.

• Journal of Genetic Medicine
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• v.1 no.1
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• pp.23-26
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• 1997

Myotonic dystrophy (DM) is caused by the expansion of CTG trinucleotide repeat near the 3' end of the gene encoding for a member of protein kinase gene family (DMPK). The normal range of the CTG repeat was determined in 178 normal individuals (141 unrelated individuals and 37 of 9 families) by polymerase chain reaction (PCR), polyacrylamide gel electrophoresis and silver staining method. And the expansion of the CTG repeats in a DM family was analyzed with Southern analysis. In normal population, the range of CTG repeat is between 5 and 34 and 19 different alleles were observed in that range, and $(CTG)_{11-14}$ alleles were predominant. 4 members of an affected family showed the 0.5-2.0 kb size expansion of CTG repeats. In this study we could predict the incidence of DM in Korea as 1 in 20,000 and we could establish the diagnostic procedure for myotonic dystrophy.

• IMMUNE NETWORK
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• v.10 no.6
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• pp.230-238
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• 2010

Background: The MK1 strain, a novel bacterial isolate from soft-rotten tissue of the Neungee mushroom, produces copious amounts of exopolysaccharide (EPS) in a dextrose minimal medium. This study examined the molecular characteristics and immunomodulatory activity of MK1 EPS. Methods: The EPS in the culture supernatant was purified by cold ethanol precipitation, and characterized by SDS- PAGE/silver staining and Bio-HPLC. The immunomodulatory activities of the EPS were examined using the mouse monocytic cell line, RAW 264.7 cells. Results: The molecular weights of the purified EPS were rather heterogeneous, ranging from 10.6 to 55 kDa. The EPS was composed of glucose, rhamnose, mannose, galactose, and glucosamine at an approximate molar ratio of 1.00 : 0.8 : 0.71 : 0.29 : 0.21. EPS activated the RAW cells to produce cytokines, such as TNF-${\alpha}$ and IL-$1{\beta}$, and nitric oxide (NO). EPS also induced the expression of co-stimulatory molecules, such as B7-1, B7-2 and ICAM-1, and increased the phagocytic activity. The macrophage-activating activity of EPS was not due to endotoxin contamination because the treatment of EPS with polymyin B did not reduce the macrophage-activating activity. Conclusion: The EPS produced from the MK1 strain exerts macrophage-activating activity.

• Journal of Oral Medicine and Pain
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• v.21 no.2
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• pp.317-329
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• 1996

Allelic frequency and genotype distribution of short tandem repeat(STR) F13A01 locus was analysed by polymerase chain reaction, polyacrylamide gel electrophoresis and silver staining from human genomic deoxyribonucleic acid(DNA) was extracted from 205 unrelated Korean to be applied to forensic identification and parentage testing as a database. The results were as follows : 1. 5 alleles and 11 genotypes of F13A01 locus were detected and heterozygosity value was 62.0% and the observed each alleles and allelic frequency was 3.2(0.363), 4(0.105), 5(0.063), 6(0.466), 16(0.002). 2. The allelic diversity value was 0.639 and the power of discrimination was 0.804.3. Compared with observed number of alleles and allele frequency in ethnic difference, result was appeared to be similar to that of Japanese and Asians, while was appeared to be much different to that of Blacks and Caucasians in the observed number of alleles and frequency of allele 3.2, 5, 7. From the above result of this investigation, the allelic frequency of STR F13A01 locus in the Korean was considerd to be useful for individual identification and parentage testing as a database.

• Journal of Microbiology
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• v.38 no.1
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• pp.11-17
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• 2000

A new method was developed for the rapid analysis of diverse bacterial species in the natural environment. Our method is based on PCR-single-strands-conformation polymorphism (PCR-SSCP) and selective isolation technique of single-stranded DNA. Variable V3 fragments of 16S rDNA were amplified by PCR with bacterial 16S rDNA primers, where one of the primers was biotinylated at the 5'-end. The biotinylated strands of the PCR products were selectively isolated by using streptavidin paramagnetic particles and a magnetic stand, to prevent SSCP analysis producing heteroduplexes from heterogeneous DNA samples. The selected strands were separated by electrophoresis on a polyacrylamide gel, and detected by silver staining. Analysis of PCR products from 8 bacterial strains demonstrated their characteristic DNA band patterns. In addition, changes in the structure of the bacterial community and species diversity in the microcosm treated with phenol could be monitored. After 3 weeks of incubation, phenol and its intermediate, 2-hydroxy-muconic-semialdehyde, were degraded by indigenous bacteria. These dominating bacterial populations were identified as strong bands on an SSCP gel. Therefore, this study provides useful tools for microbial community analysis of natural habitats.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.12
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• pp.1685-1690
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• 2006

Kenkatha cattle, a draft purpose breed, which can survive in a harsh environment on low quality forage, was explored genetically exploiting FAO-suggested microsatellite markers. The microsatellite genotypes were derived by means of the polymerase chain reaction (PCR) followed by electrophoretic separation in agarose gels. The PCR amplicons were visualized by silver staining. The allelic as well as genotypic frequencies, heterozygosities and gene diversity were estimated using standard techniques. A total of 125 alleles was distinguished by the 21 microsatellite markers investigated. All the microsatellites were highly polymorphic with mean allelic number of 5.95${\pm}$1.9 (ranging from 3-10 per locus). The observed heterozygosity in the population ranged between 0.250 and 0.826 with a mean of 0.540${\pm}$0.171, signifying considerable genetic variation. Bottleneck was examined assuming all three mutation models which showed that the population has not experienced bottleneck in recent past. The population displayed a heterozygote deficit of 21.4%. The study suggests that the breed needs to be conserved by providing purebred animals in the breeding tract.

• KSBB Journal
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• v.9 no.2
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• pp.115-121
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• 1994

A Purification device with 30-channel free-flow electrophoresis was assembled to treat samples of 240m1 volume for purification of lentil lectin (LcH) from lentil seeds with no impurities in a silverstained PAGIEF gel. HEPES(50mM)-Ttis(50mM), Cycloserine(50mM)-urea(3M), Histidine(50mM)-urea(3M) were used as ampholytea among which Histidine(50mM)-urea(3M) (pI 7.65) was found best in resolution. LcH is known to be present in the form of LcH-A, LcH-B and the complex of the two. The complex, however, disappeared when urea was added in the ampholytes, which suggested that the complete purification of two isolectins is possible using the present multistep purificaton device.