• Journal of Microbiology and Biotechnology
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• v.10 no.6
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• pp.805-810
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• 2000

An antifungal protein antagonistic to the rice blast fungus, Pyricularia oryzae was purified from Paenibacillus macerans PM-1 by ammonium sulfate fractionation, Q Sepharose Fast Flow column chromatography, Phenyl Sepharose CL-4B column chromatography and Superose 12 gen filtration. An apparent molecular mass of the purified antifungal protein was determined as 8 kDa by SDS-PAGE and 9 kDa by analytical gel filtration, respectively, suggesting that the purified protein is a monomer. The antifungal protein was stable at pH range from 7-12 and up to $100^{\circ}C$. The protein was also stable at 0.1-1% Tween 20 and Triton X-100. The N-terminal amino acid sequence of the antifungal protein was Thr-Glu-Leu-Pro-Leu-Gly-Ile-Val-Met-Asp-Lys-Tyr-Thr-Asp-Ala-Phe-Lys-Phe-Asp-Met-Phe. Comparison of the determined sequence with other peptide and DNA sequences did not reveal homology at all. Therefore, the purified antifungal protein was speculated to be a novel protein. The condidial germination in vitro of P. oryzae KJ301:93-39 by the purified protein ($5.9{\mu} g/ml$) was limited to $9{\pm}3.2%$ only, compared with $69{\pm}2.4%$ of the control. Ungerminated conidia were swollen at basa and mid cell by the purified protein. In vivo bioassay for inhibition of conidial germination of P. oryzae KJ 301, one of the most predominating racesin Korea. the purified protein ($5.9{\mu} g/ml$)strongly inhibited the conidial germination. The conidia, even though germinated, could not develop any further to produce appressoria efficiently.

• Korean Journal of Microbiology
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• v.23 no.2
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• pp.115-121
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• 1985

Five representative virulent phages (J1, TK93, K1, PD5, and CP1) and one temperate phage (.phi.1043) of Lactobacillus casei were compared to each other by analyzing the agarose gel electrophoretic patterns of restriction enzyme-digested phage DNAs. Nucleic acids of all the tested phages were double stranded DNA. DNAs of J1, TK93, K1, and ${\phi}$ 1043 phages had a size of about 42kb, but the size of PD5 and CP1 DNAs was avout 140kb. J1, TK93, K1, PD5, CP1, and ${\phi}$ 1043 DNAs were digested to 13, 13, 11, 14, 14, and 12 fragments by EcoR1, respectively, and showed its characteristec restriction patterns. Cohesive ends were present in J1, TK93, and ${\phi}$ 1043, but were absent in K1, PD5, and CP1. Restriction maps of J1 and TK93 DNAs showed nearly complete homology and their evolutionary relationship based upon the restriction analysis was discussed.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.120-120
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• 2017

Since the composition of proteins from the Korean cultivars of Proso millet is unknown; thereby, the present study was conducted to obtain a reference map of millet seed proteins and identify the functional characteristics of the identified proteins. Proteins extracted from the millet seeds of various cultivars, were investigated using proteomic techniques as 2D electrophoresis coupled with mass fingerprinting. The 1152 (differentially expressed) proteins were detected on 2-D gel. Among them, 26 reproducible protein spots were analyzed by MALDI-TOF-TOF mass spectrometry. Out of 26 proteins, 2 proteins were up-regulated towards all cultivars of millet, while 7 proteins were up-regulated and 13 proteins were down-regulated against only one cultivar. However, abundance in most identified protein species, associated with metabolism, transcription and transcription was significantly enhanced, while that of another protein species involved in polysaccharide metabolism, stress response and pathogenesis were severely reduced. Taken together, the results observed from the study suggest that the differential expression of proteins from the four cultivars of millet may be cultivar-specific. Taken together, a proteomic investigation of millet seeds from different cultivars, we sought to better understand the genetic variation of millet cultivars representing the future millet research, and the functional categorization of individual proteins on the basis of their molecular function.

• Mycobiology
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• v.44 no.4
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• pp.283-290
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• 2016

A double-stranded RNA (dsRNA) mycovirus was detected in malformed fruiting bodies of Pleurotus ostreatus strain ASI2792, one of bottle cultivated commercial strains of the edible oyster mushroom. The partial RNA-dependent RNA polymerase (RdRp) gene of the P. ostreatus ASI2792 mycovirus (PoV-ASI2792) was cloned, and a cDNA sequences alignment revealed that the sequence was identical to the RdRp gene of a known PoSV found in the P. ostreatus strain. To investigate the symptoms of PoV-ASI2792 infection by comparing the isogenic virus-free P. ostreatus strains with a virus-infected strain, isogenic virus-cured P. ostreatus strains were obtained by the mycelial fragmentation method for virus curing. The absence of virus was verified with gel electrophoresis after dsRNA-specific virus purification and Northern blot analysis using a partial RdRp cDNA of PoV-ASI2792. The growth rate and mycelial dry weight of virus-infected P. ostreatus strain with PoV-ASI2792 mycovirus were compared to those of three virus-free isogenic strains on 10 different media. The virus-cured strains showed distinctly higher mycelial growth rates and dry weights on all kinds of experimental culture media, with at least a 2.2-fold higher mycelial growth rate on mushroom complete media (MCM) and Hamada media, and a 2.7-fold higher mycelial dry weight on MCM and yeast-malt-glucose agar media than those of the virus-infected strain. These results suggest that the infection of PoV mycovirus has a deleterious effect on the vegetative growth of P. ostreatus.

• The Plant Pathology Journal
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• v.22 no.3
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• pp.260-264
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• 2006

Talaromyces flavus mediates the transmission of Apple stem grooving virus (ASGV) to several host plants. The ASGV-F carried by T.flavus was partially purified from the fungus. Based on sequence analysis and homology searches, this is closely related to other ASGV strains isolated from host plants. The partially purified viral coat protein (CP) was separated on a 12% SDS-polyacrylamide gel and analyzed by Western blotting with an ASGV anti-serum. A single band at 28 kDa reacted with the ASGV anti-serum. The deduced amino acid sequence of the ORF-l showed conserved domains, including an NTP-binding helicase motif, GFAGSGKT. The amino acid sequences of the helicase and CP showed strong homology to other ASGV strains (98%). All ASGV isolated from plants and fungi had salt bridges composed of the CP and the GFAGSGKT motif of the helicase, which are commonly conserved in plant viruses. These results suggest that ASGV-F is one of ASGV strains isolated from T.flavus based on sequence similarity as well as the serological analysis of CP.

• Korean Journal of Medicinal Crop Science
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• v.12 no.5
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• pp.349-354
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• 2004

The extracts of Pinus densiflora sawdust by n hexane, ethyl acetate and methanol solvent were investigated to identify their mycelial growth inhibition against Lentinus edodes. The yields of n hexane soluble fraction, ethyl acetate-soluble fraction, and methanol soluble fraction from P. densiflora sawdust were obtained 1.36%, 2.21% and 4.03% using organic solvent, respectively. The mycelial growth inhibition of L. edodes was the greatest for n hexane extract, ranging from 36.5% to 47.6% at concentrations of 125 ppm to 1,000 ppm, with the values for all concentrations significantly different from one another. After direct extraction of P. densiflora sawdust using n hexane, ethyl acetate and methanol, each extract was separated into three fractions by silica gel column chromatography and then the fractions were isolated on the values of $R_f$ by thin layer chromatography. The mycelial growth inhibition against L. edodes was recognized in the fractions II (33.5%) and III (37.6%) of n hexane extract, the fraction II (21.4%) of ethyl acetate extract and the fraction II (26.4%) of methanol extract. The fractions III of n-hexane extract showed the highest growth inhibition among the nine fractions of the organic solvent extract.

• Korean Journal of Veterinary Research
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• v.29 no.4
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• pp.445-455
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• 1989

The proteins of the ruminant erythrocyte membranes were analysed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate, and their relations to the slow erythrocyte sedimentation rate(ESR) of the ruminants were investigated by treating the erythrocytes with proteinases such as trypsin, chymotrypsin and pronase, and glycosidases such as neuraminidase and galactosidase. Protein content in the erythrocyte membrane was $2.85{\pm}0.28$ in human, $3.60{\pm}0.41$ in Korean cattle, $3.71{\pm}0.36$ in Holstein, $4.13{\pm}0.83$ in Korean native goat and $3.94{\pm}0.56mg/ml$ in sheep, showing higher in ruminant animals than in human(p<0.01). Although the general protein profiles of the ruminant erythrocyte membranes were almost similar to that of human, all the ruminant erythrocyte membranes showed one additional protein band, called band-Q in the previous report on proteins of bovine erythrocyte membrane, which migrated electrophoretically to the mid position between band-2 and band-3 in human erythrocyte membranes. The glycoprotein profiles of ruminant erythrocyte membranes revealed by periodic acid Schiff(PAS) stain showed a marked difference from that of human. The PAS-1(glycophorin) and PAS-2(sialoglycogrotein) present in human erythrocyte membranes were almost absent from the ruminant animals. Instead, a strong PAS-positive band near the origin of the electrophorograms, which was named as PAS-B in the previous report on proteins of bovine erythrocyte membranes, was shown in the ruminant animals except sheep. In addition, the erythrocyte membranes of Korean native goat and sheep showed a moderate PAS-negative band near the tracking dye of the electrophorograms, which was named as PAS-G in this study. In the erythrocyte treated with the enzymes, the migration of each protein fracture of erythrocyte membranes in response to each enzyme was diverse according to different species or breed of ruminant animals. Among others, band-Q present in ruminants was slightly or moderately decreased by trypsin-, chymotrypsin-, and pronase- treatments of the erythrocytes, but not only in sheep. It was particularly noticeable that PAS-B, a fraction of glycoprotein, present in ruminants except sheep, was better digested by proteinases than by glycosidases, showing remarkable increase(p<0.01) of the ESR in accord with complete digestion(disappearance) of the PAS-B band by pronase, trypsin or chymotrypsin treatment of erythrocytes. In sheep, there was almost no any response to the various enzymes in general protein and glycoprotein profiles of the erythrocyte membranes except PAS-G, which was markedly decreased by pronase treatment of the erythrocytes. Nevertheless, the ESRs were accelerated in erythrocytes treated with pronase, trypsin, chymotrypsin and neuraminidase. Erythrocyte osmotic fragility was increased in erythrocytes treated with only pronase among five enzymes in all the human and ruminant animals used in this study.

• Restorative Dentistry and Endodontics
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• v.15 no.1
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• pp.107-119
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• 1990

The purpose of this study was to evaluate the marginal adaptability of the amalgam restorations in applying the cavity varnish (Copalite$^{(R)}$) and dentin bonding agent (Scotchbond 2$^{(R)}$) under the scanning electron microscope. For this study, eighteen sound extracted human molars were selected. Class I cavities in 12 teeth and class V cavities in 6 teeth were prepared using an air turbine with No. 701 tungsten carbide bur and finished using a low speed handpiece with No. 557 fissure bur. The prepared specimens were then divided into three groups including 4 class I cavities and 2 class V cavities in each group and restored as follows ; Group I. All the prepared cavities were restored with amalgam only (Control). Group II. Two layers of Copalite$^{(R)}$ cavity varnish were applied to the cavities with a gentle stream of air after each application and cavities were restored with amalgam. Group III. The enamel cavity margins were etched with 37% phosphoric acid gel for 60 sec., rinsed for 30 sec. and dried. One layer of visible lightcured Scotchbond Dental Adhesive$^{(R)}$ was applied and immediately cured for 20 seconds with visible light-cure unit and cavities were restored with amalgam. All the specimens were cut at the neck of the teeth and the occlusal halves of specimens were sectioned buccolingually in the longitudinal axis centering the amalgam restorations, using the disk. The cut specimens were ground with sandpapers (400, 600, 800, 1000 grit), and cleaned for 5 minutes in the ultrasonic cleaner (Brason Co. U.S.A.). In the cut surfaces, the amalgam - tooth interfaces were examined under the scanning electron microscope (JSM, 35C type, JEOL). The obtained results were as follows ; 1. The amalgam-tooth interfaces were reduced more significantly in the Copalite$^{(R)}$ and Scotchbond 2$^{(R)}$ application group than in the control group. 2. In the class I cavities, the Scotchbond 2$^{(R)}$ application group showed the findings similar to the Copalite$^{(R)}$ application group in the cavity floor, and the marginal adaptability was better in the side wall than in the cavity floor. 3. In the class I cavities, the Scotchbond 2$^{(R)}$ application group showed better marginal adaptability in the occlusal margin than in the gingival margin. 4. The marginal adaptability was in the order of the Scothbond 2$^{(R)}$ application group, the Copalite$^{(R)}$ application group and the control group.

• Korean Journal of Microbiology
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• v.44 no.4
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• pp.305-310
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• 2008

Staphylococcus aureus is one of the most significant pathogens and a causative agents of nosocomial infections. The emergence of methicillin resistant S. aureus (MRSA), in particular, has become a major clinical and epidemiological problems worldwide. In this study, we analyzed the toxin genes and investigated molecular epidemiological characteristics of S. aureus isolated from stools of diarrheal patients at the hospitals in Incheon. Of the 609 strains from 2,281 specimens, 173 strains retained enterotoxin; 68 isolates (39.30%), 100 isolates (57.80%) were classified to A and C type, respectively. In the antibiotic susceptibility, all of enterotoxin positive isolates were resistant to oxacillin. Eighty eight strains (50.86%) of 173 MRSA isolate possessed tsst gene, but eta and eth genes were not detected at all. In the detection of MRSA associated genes by PCR method, mecA genes were detected in 167 strains (96.53%). From the result of PFGE analysis, we classified tsst-positive MRSA to 10 types and 24 subtypes. Type A, H and F were the major strains comprised of 57.95% (51 strains), 10.22% (9 strains) and 9.09% (8 strains) respectively.

• Journal of Periodontal and Implant Science
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• v.29 no.1
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• pp.1-14
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• 1999

Ursodeoxycholic acid(UDCA) is a hydrophilic gall bladder acid and has been used as a effective drug for liver disease related to in1munity. This drug inhibits secretions of IL-2, IL-4, and $IFN-{\gamma}$ from T-cells and production of immunoglobulin from B-cells. Also it has been reported that UDCA inhibits production of IL-1 related to the progression of periodontal disease and activation of collagenases. The purpose of the present study was to elucidate the effects of UDCA on inhibition of periodontal disease progression using clinical, microbiological and histometrical parameters. Twelve pure bred, 16 month-old-beagle dogs were used in the study. After ligature-induced periodontal diseases were formed, experimental drugs were applied twice a day and then the results of clinical, microbiological, and histometrical parameters were measured at baselie(initiation of experiment) , 4weeks and 8weeks. The gel with UDCA(concentration 0.5%, 5% 3 dogs in each) was applied to experimental group, chlorhexidine to positive control group(3dogs) and the gel without UDCA(base) to negative control group. After induction of general anesthesia, the maxillary 2nd, 3rd premolars and 1st molar and the mandibular 2nd, 3rd, 4th premolars and 1st molar were ligated in one side selected randomly and were not ligated in the opposite side. The plaque index(PI), gingival index(GI), pocket depth(PD) and gingival crevicular fluid(GCF) volum were measured clinically. The PI and GI were measured at 3 buccal points of all experimental teeth and the GCF was measured only at the 3rd premolar in the maxilla and the 4th premolar in the mandible. In the microbiological study, the samples extracted from the 3rd premolar of the maxilla and the 4th premolar of the mandible at the center of buccal surface were analyzed aerobics, anaerobics and Streptococcus colony forming units, After clinical and microbiological examination at 8weeks, the dogs were sacrificed by carotid artery perfusion. The samples were fixed and sectioned including interproximal area, and the distance from cementoenamel junction(CEJ) to alveolar crest was measured. The results were that PI, GI and PD increased until 4 weeks and decreased at 8 weeks in three groups but the differences between all the groups were not significant. The 0.5% UDCA in non-ligated group showed remarkable decrease of GCF. The experimental group applied 5% UDCA decreased the number of aerobics and anaerobics. The distance from CEJ to alveolar crest was greater in the negative control group on both ligated and non-ligated sides, but the differences were not significant stastically.