• Food Science and Biotechnology
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• 제17권4호
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• pp.761-765
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• 2008

Bacillus cereus causes two different types of food poisoning syndromes: diarrhea and emesis. The diarrheal syndrome is attributed to various enterotoxins, including nonhemolytic enterotoxin, hemolytic enterotoxin, and enterotoxin-T, whereas the emetic syndrome is caused by the dodecadepsipeptide toxin cereulide. A multiplex polymerase chain reaction (PCR) assay was developed to rapidly detect and identify B. cereus strains. Three primer pairs specific to regions within genes encoding nonhemolytic enterotoxin (nheA), molecular chaperonin (groEL), and cereulide synthetase (ces) were used to identify and differentiate between the enterotoxin-producing and emetic toxin-producing B. cereus strains. The cereulide-producing emetic B. cereus showed 3 PCR products of 325, 405, and 685 bp for the groEL, ces, and nheA genes, respectively, whereas the enterotoxin-producing B. cereus showed 2 PCR products without a ces gene specific DNA fragment. Specific amplifications and differentiations by multiplex PCR assay were obtained using 62 B. cereus strains and 13 strains' of other bacterial species. The detection limit of this assay for enterotoxin-producing strain and emetic toxin-producing strain from pure cultures were $2.4{\times}10^1$ and $6.0{\times}10^2\;CFU/tube$, respectively. These results suggest that our multiplex PCR method may be useful for the rapid detection and differentiation of B. cereus strains in foods.

• 한국버섯학회지
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• 제16권2호
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• pp.103-110
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• 2018

11 종의 목이버섯 자실체로부터 얻은 50% EtOH추출물을 사용하여 지방전구세포인 3T3-L1의 지방세포 분화억제 효과를 확인한 모든 strain에서 억제효과가 존재하였다. 털목이에서는 21001 균주가 가장 우수하였고 흑목이에서는 21002 균주가 억제효과가 우수하였다. 전반적으로 털목이가 흑목이에 비해 지방세포 분화억제 효과가 우수하였다. 또한 Adipocyte RNA 발현량과 protein 발현량 측정 실험에서도 털목이 21001 균주가 지방세포 분화억제 효과에 있어서 가장 우수하였다. 털목이 21001 버섯을 사용하여 동물실험을 진행한 결과 털목이 자실체 분말이나 50% EtOH 추출물을 급여한 실험에서 체중 감소효과가 나타났으며, 특히 EtOH 추출물 0.2%를 급여한 군에서 유의하게 감소함을 확인하였다(*P<0.05). 동물 희생 후 적출한 간 중량 및 부고환지방량과 혈청 내 중성지방 및 총 콜레스테롤, 혈당, ALT, AST 등도 털목이버섯 21001 EtOH 추출물 0.1% 이상 급여 시 고지방식이만을 급여한 군에 비해 유의차(*P<0.05 이상) 있게 감소함을 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제9권6호
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• pp.744-750
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• 1999

Forty alcohol yeast strains were isolated from the main mashes (10 strains from each mash) for brewing of 4 different kinds of Korean traditional liquor (3 different types of Yakju and 1 Takju). Thirty-eight out of 40 strains were identified to be the same strain, Saccharomyces boulardii, by the Automated Bacteria, Yeast, and Fungi Identification System (Biolog Co., U.S.A.) based on the metabolic fingerprints. One strain that showed the highest ethanol production among the 38 strains in YPD medium, designated SHY 111, was selected and used for differentiating from other yeast type strains using the polymerase chain reaction (PCR). Amplified DNA, from transcribed internal spacers of SHY 111 chromosomal DNA, was found to be the same in both size and sequence as those of S. cerevisiae KCCM 11215 (formerly S. coreanus) and S. boulardii along with that of S. cerevisiae AB 972, which was used as a type strain for the yeast genome project. However, when PCR was carried out with the intron splice site primer, it resulted in the amplification of the SHY 111-specific DNA fragment which was about 200 bp in size. When PCR was carried out using the primer to test diversity of 40 isolated yeast strains, it was found that the PCR patterns were similar to each other except for the 200 bp bands derived from all the 10 strains from one Yakju, and 2 strains from another Yakju. These results suggest the strain identified as S. boulardii by the Automated Identification System to be a dominant strain for the fermentation of Korean traditional liquors.

• 한국미생물·생명공학회지
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• 제22권5호
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• pp.467-476
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• 1994

Lactobacillus casei HY 2782, prophage cured strain was characterized to be stable as much as L casei YIT 9018, parent strain. By southern hybridization, it was confirmed that the temperate phage was incorporated in chromosomal DNA of L. casei YIT 9018 as a prophage. It was also proved that the prophage was cured from chromosomal DNA of L casei HY 2782. The growth rate, lactic acid producing ability, carbohydrates fermentation, and enzymatic activity of L. casei HY 2782 were found to be similar to those of L. casei YIT 9018. When L casei HY 2782 was used as a host, the multiplicity of infection (M.O.I.) of the temperate phage for L. casei HY 2782 was 1.0~5.0. Restriction enzyme analysis of pLC90 plasmid from L. casei HY 2782 was shown that the size was an approximately 68.22 kb. The plasmid profiles, genomic DNA patterns, and cellular fatty acids composition of L. casei HY 2782 were similar to those of L casei YIT 9018. And the major fatty acids composition of these strains were C$_{14;0}$,C$_{16;1}$, C$_{16;0}$, C$_{18;1}$ and C$_{19;cyclo-}$ 10 sets of arbitrary primer in the PCR were screened to find differentiation against two strains of L. casei. Among them, b$_{5}-1/17-1 primer was produced an approximately 1.3 kb DNA band of only L casei YIT 9018. And b$_{5}-2/17-2 primer was produced an approximately 1.0 kb DNA band of only L casei HY 2782.

• 대한수의학회지
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• 제39권4호
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• pp.778-785
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• 1999

To detect CDV RNA in clinical samples and differentiate prevailing CDV virulent strains affecting susceptible animals from attenuated vaccine strains, we performed RT-PCR, RFLP, and sequencing. CDV specific primers were generated from the middle part of nucleocapsid gene. The expected size of PCR products, 519 bp, was observed in tissues of Jindo dog, poodle dog, badger, fourteen of nineteen blood samples as well as 5 vaccine strains including domestic and imported products. The PCR products obtained from tissues and PBMCs of infected animals were digested to 317- and 202-bp fragments by Bam HI, but the products obtained from four of five vaccine strains and Lederle strain were not digesed by Bam HI. Only one vaccine strain of which the PCR products were digested by Bam HI was confirmed as imported vaccine, modified Synider Hill strain. Based on seqencing data obtained from the 519-bp products, it was confirmed that Bam HI restriction site tends to be conserved in field isolates compared to the commercially available attenuated vaccine strains. Partial nucleotide sequences of CDV NP gene obtained from tissues of Jindo dog, poodle and badger shared 100% homology each other, whereas the nucleotide sequences showed 96.3, 96.5, 93.6 and 93.4% homology with Yanaka (virulent), Han95 (virulent), Lederle (attenuated) and Onderstepoort (attenuated) strain, respectively.

• Mycobiology
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• 제46권4호
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• pp.382-387
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• 2018

The entomopathogenic fungus Cordyceps militaris is a valuable medicinal ascomycete, which degenerates frequently during subsequent culture. To avoid economic losses during industrialized production, scratching stimuli of mycelia was introduced to improve the fruiting body production. The present results indicated that higher yields and biological efficiency were obtained from two degenerate strains (YN1-14 and YN2-7) but not from g38 (an insertional mutant in Rhf1 gene with higher yields and shorter growth periods). Furthermore, the growth periods of the fruiting bodies were at least 5 days earlier when the mycelia were scratched before stromata differentiation. Three ROS-scavenging genes including Cu/Zn superoxide dismutase (CmSod1), Glutathione peroxidase (CmGpx), and Catalase A (CmCat A) were isolated and their expression profiles against scratching were determined in degenerate strain YN1-14 and mutant strain g38. At day 5 after scratching, the expression level of CmGpx significantly decreased for strain g38, but that of CmSod1 significantly increased for YN1-14. These results indicated that scratching is an effective way to promote fruiting body production of degenerate strain, which may be related at least with Rhf1 and active oxygen scavenging genes.

• 한국정보통신학회:학술대회논문집
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• 한국정보통신학회 2013년도 춘계학술대회
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• pp.1012-1014
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• 2013

DNA-DNA hybridization method with four oligonucleotide-specific probes was used simultaneously for differentiation and identification of four Mycobacterium species (Mycobacterium tuberculosis, M. avium, M. intracellulare, and M. kansasii). This DNA-DNA hybridization method with 4 oligonucleotide-specific probes, which targets in the rpoB region of 4 Mycobacteria species, respectively, was tested on 322 clinical isolates. Using DNA-DNA hybridization method, we detected M. tuberculosis (282 strains), M. avim (7 strains), M. intracellulare (9 strains), and M. kansasii (3 strain) from 322 clinical isolates. This result was compared with conventional biochemical test and rpoB DNA sequence analysis of this clinical isolates. We confirmed identification of Mycobacterium tuberculosis, M. avium, M. intracellulare, and M. kansasii with high sensitivity (100 %) and specificity (100 %). This DNA-DNA hybridization method could be performed within 4 hours at least. Therefore, we suggest that DNA- DNA hybridization method using 4 rpoB DNA probes of Mycobacteria could be used for accurate, rapid, convenient detection and identification of Mycobacterium tuberculosis, M. avium, M. intracellulare, and M. kansasii in clinical samples.

• 한국식물병리학회지
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• 제12권1호
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• pp.11-20
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• 1996

한 쌍의 R16-1과 R23-2R primer를 이용한 PCR에 의해 증폭된 16S와 23S rDNA 사이의 rDNA spacer 부위의 다형성들이 Pseudomonas avenae, P. glumae, P. fuscovaginae, P. syringae pv. syrngae, Xanthomonas oryzae pv. oryzae, X. oryzae, Xanthomonas herbicola 등 벼 종자전염성 51개 균주의 구분을 위하여 적용되었다. 증폭산물은 820∼950bp의 크기였으며, 각각의 종에 특이적이었고 구분이 가능하였다. Pseudomonas species의 증폭산물은 P. avenae는 950bp, P. glumae는 850bp, P. fuscovaginae는 770pb 및 P. syringae pv. syringae는 1,240, 1,100 및 820bp로 특이적이었다. P. avenae와 P. glumae의 국내균주들은 다형성에 있어 종내 변이는 없었다. X. oryzae pv. oryzae의 860bp와 X. oryzae pv. oryzicola의 890, 440 및 370bp의 이차산물에서 Xanthomonas species의 종내에서 균주에 관련없이 단일화된 다형성을 보였다. CXO 211을 제외한 모든 국내 균주는 a형에 속한 반면 하나의 국내 균주를 포함하여 4개 균주는 b형이었다. E. herbicola의 spacer 부위 증폭은 여러 개의 band를 보였으며, 증폭상은 각각 동일하였고, strain간의 종내 변이는 없었다. 본 실험 결과에 의하여 16S와 23S rDNAdp R16-1과 R23-2R primer를 이용하여 PCR 증폭된 spacer 다형성의 구별은 종자전염성 세균의 신속한 구별에 이용될 수 있을것이다.

• 식물병연구
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• 제18권2호
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• pp.129-132
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• 2012

Citrus bacterial canker is an economically important disease affecting citrus production in many citrusgrowing areas and several pathotypes have been recognized within the Xanthomonas pathogens causing canker. In view of the containment of the disease, accurate identification of the causal bacterium is important. In this study, triplex PCR method was developed by using the previously reported primers. Two groups of primer combination, such as, one group including primers 2/3, J-pth1/J-pth2 and XACF/XACR, and another group 2/3, J-pth1/J-pth2 and Xac01/Xac02, were suitable for the detection and differentiation of X. a. pv. citri $A^w$, X. a. pv. aurantifolii B and C, and X. a. pv. citrumelo E strains. Moreover, the primer combination of Xac01 and J-pth2 promised us to use as a specific primer set to detect X. a. pv. citrumelo E strain. The PCR methods developed in this study could be used for the rapid differentiation of Xanthomonas pathotypes of citrus.

• Journal of Microbiology and Biotechnology
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• 제17권5호
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• pp.705-711
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• 2007

Two novel strains of the Cytophaga-Flexibacter-Bacteroides(CFB) group, designated Gsoil $219^T$ and Gsoil $238^T$, were isolated from soil of a ginseng field of Pocheon Province in Korea. Both strains were Gram-negative, aerobic, nonmotile, nonspore-forming, and rod-shaped. Phylogenetic analysis based on 16S rRNA gene sequences indicated that both isolates belong to the genus Chitinophaga but were clearly separated from established species of this genus. The sequence similarities between strain Gsoil $219^T$ and type strains of the established species and between strain Gsoil $238^T$ and type strains of the established species ranged from 91.4 to 94.7% and 91.6 to 94.2%, respectively. Phenotypic and chemotaxonomic data(major menaquinone, MK-7; major fatty acids, $iso-C_{15:0}\;and\;C_{16:1}\omega5c$; major hydroxy fatty acid, $iso-C_{17:0}3-OH$; major polyamine, homospermidine) supported the affiliation of both strains Gsoil $219^T$ and Gsoil $238^T$ to the genus Chitinophaga. Furthermore, the results of physiological and biochemical tests allowed genotypic and phenotypic differentiation of both strains from the other validated Chitinophaga species. Therefore, the two isolates represent two novel species, for which the name Chitinophaga soli sp. nov.(type strain, Gsoil $219^T=KCTC\;12650^T=DSM\;18093^T$) and Chitinophaga terrae sp. nov.(type strain, Gsoil $238^T=KCTC\;12651^T=DSM\;18078^T$) are proposed.