• Journal of Microbiology and Biotechnology
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• v.27 no.11
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• pp.2052-2059
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• 2017

The emergence and dissemination of Salmonella genomic island 1 (SGI1) are strongly associated with the occurrence of multidrug-resistant (MDR) enterobacteria in humans and animals. Diverse SGI1s have been reported among Salmonella enterica and Proteus mirabilis in several countries. We aimed to characterize SGI1 in P. mirabilis isolates from humans and chickens in Chungcheong Province, Korea. A total of 44 P. mirabilis isolates were recovered from humans (n = 20) and chickens (n = 24). Antimicrobial susceptibility was determined by disk diffusion assay. To detect and characterize SGI1s, PCR amplification and PCR mapping experiments were performed. Repetitive extragenic palindromic-PCR (REP-PCR) was performed to assess the clonality of the isolates. The four P. mirabilis strains (16.7%) from chicken harbored a SGI1, whereas none of the isolates from clinical specimens contained SGI1. The SGI1s detected in our study were all confirmed as SGI1-PmABB harboring aminoglycoside-resistant genes (aacCA5 and aadA7). In P. mirabilis isolates, the presence of SGI1-PmABB was significantly correlated with high resistance rates of the isolates to antimicrobial agents, such as gentamicin, streptomycin, and spectinomycin. Moreover, the four SGI1-bearing isolates showed the same REP-PCR patterns and that suggested both horizontal and clonal spread of the isolates. This study is the first attempt to determine SGI1s in P. mirabilis isolates in Korea. We confirmed that P. mirabilis isolates carrying SGI1-PmABB were distributed at poultry farms in Korea. The present study emphasizes the need for continuous monitoring of SGI1s to prevent spreading of the MDR genomic islands among P. mirabilis isolates from humans and animals.

• Research in Plant Disease
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• v.13 no.1
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• pp.24-29
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• 2007

A total of 35 strains of Ralstonia solanacearum isolated from wilted pepper and tomato plants in Korea were analyzed for their genetic diversity by bacteriological, pathological and molecular biological approaches. All the strains were identified as R. solanacearum biovar 4 on the basis of physiological and biochemical tests, and species-specific PCR primers. Pathogenicity of the strains was confirmed by inoculating on 4-week-old pepper and tomato seedlings. Using cluster analysis based on repetitive sequence-based polymerase chain reaction (rep-PCR) genomic fingerprints, R. solanacearum strains isolated from pepper and tomato in Korea divided into 6 groups showing a high degree of genetic diversity at 55% similarity level. The genetic diversify of strains was not significantly correlated with their geographic origins and host plants.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.312-320
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• 2000

Abstract The full sequence of the plasmid pKJ36, which was derived from Bifidobacterium longum KJ, was determined and analyzed to construct shuttle vectors between E. coli and Bifidobacterium. The plasmid pKJ36 was composed of 3,625 base pairs with a 65.1% G+C content. The structural organization of pKJ36 was highly similar to that of pKJ50, and the three major ORFs on pKJ36 showed high amino acid sequence homologies with those of pKJ50. The putative proteins coded by these three ORFs were designated as RepB (32.0 kDa, pI=9.25), MembB (29.0 kDa, pI=12.25), and MobB (39.0 kDa, pI=IO.66), respectively. The amino acid sequence of RepB showed a 57% identity and 70% similarity with that of the RepA protein of pKJ50. Upstream of the repB gene, the so-called iteron sequence was directly repeated four-and-ahalf times and a conserved dnaA box was identified. An amino acid sequence comparison between the MobB and MobA of pKJ50 revealed a 48% identity and 61 % similarity. A conserved oriT sequence with an inverted repeat identical to that of pKJ50 was also found upstream of the mobB gene. A hydropathy analysis of MembB revealed four possible transmembrane regions. The expressions of the repB and membB genes were confirmed by RT-PCR. The in vitro translation reaction of pKJ36 showed protein bands with anticipated sizes with respect to each putative gene product. S 1 endonuclease treatment and Southern hybridization suggested that pKJ36 replicates by a rolling circle mechanism via a single-stranded DNA (ssDNA) intermediate. A shuttle vector between E. coli and Bifidobacterium sp. was constructed using the pKJ36, pBR322, and staphylococcal chloramphenicol acetyl transferase (CAT) gene. The successful transformation of the Bifidobacterium strains was shown by Southern hybridization and PCR. The transformation efficiency differed from strain to strain and, depending on the electroporation conditions, with a range between $1.2{\times}10^1-2.6{\times}10^2{\;}cfu/\mu\textrm{g}$ DNA.X> DNA.

• The Plant Pathology Journal
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• v.27 no.4
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• pp.360-366
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• 2011

Bacterial leaf spot of Euphorbia pulcherrima has been reported in many countries. Characterization by polyphasic approaches indicated that the isolates from India, USA and New Zealand could be distinguished based on rep-PCR profiles and gyrB phylogenies, while the Chinese isolates should be ascribed to Xanthomonas axonopodis pv. poinsettiicola.

• The Plant Pathology Journal
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• v.26 no.3
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• pp.223-230
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• 2010

The Burkholderia cepacia complex isolates causing bacterial fruit rot of apricot were characterized by speciesspecific PCR tests, recA-HaeIII restriction fragment length polymorphism (RFLP) assays, rep-PCR genomic fingerprinting, recA gene sequencing, and multilocus sequence typing (MLST) analysis. Results indicated that the isolates Bca 0901 and Bca 0902 gave positive amplifications with primers specific for B. vietnamiensis while the two bacterial isolates showed different recA-RFLP and rep-PCR profiles from those of B. vietnamiensis strains. In addition, the two bacterial isolates had a higher proteolytic activity compared with that of the non-pathogenic B. vietnamiensis strains while no cblA and esmR marker genes were detected for the two bacterial isolates and B. vietnamiensis strains. The two bacterial isolates were identified as Burkholderia seminalis based on recA gene sequence analysis and MLST analysis. Overall, this is the first characterization of B. seminalis that cause bacterial fruit rot of apricot.

• YAKHAK HOEJI
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• v.57 no.2
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• pp.132-138
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• 2013

Acinetobacter baumannii is gram-negative bacilli that can be widely found in environments. Recently, A. baumannii emerged as a serious nosocomial infection. A total of 92 A. baumannii were isolated from hospitalized patients in Seoul, Korea, between December 2010 and April 2011. Antimicrobial susceptibility testing was investigated using CLSI agar dilution methods. Tigecycline non-susceptible A. baumannii isolates were investigated by repetitive extragenic palindromic sequence-based PCR (rep-PCR). Pulsed-field gel electrophoresis was performed to determine the epidemiological relationships. All clinical isolates showed high-level resistance to the most commonly used antibiotics: Ciprofloxacin (87.0%), Ampicillin/sulbactam (82.6%), Cefotaxime (81.5%), Ceftazidime (80.4%). Moreover, 50.0% of these isolates were non-susceptible to tigecycline. When evaluated by RAPD analysis, generated distinct band ranging in size from 1kb to 8k band varying from 4 to 10 bands. Stricter surveillance and more rapid detection are essential to prevent the spread of multi drug resistant A. baumannii.

• Journal of Microbiology and Biotechnology
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• v.26 no.8
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• pp.1481-1489
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• 2016

The emergence and dissemination of carbapenemase-producing Acinetobacter baumannii isolates have been reported worldwide, and A. baumannii isolates harboring bla_OXA-23 are often resistant to various antimicrobial agents. Antimicrobial resistance can be particularly strong for biofilm-forming A. baumannii isolates. We investigated the genetic basis for carbapenem resistance and biofilm-forming ability of multidrug-resistant (MDR) clinical isolates. Ninety-two MDR A. baumannii isolates were collected from one university hospital located in the Chungcheong area of Korea over a 5-year period. Multiplex PCR and DNA sequencing were performed to characterize carbapenemase and bap genes. Clonal characteristics were analyzed using REP-PCR. In addition, imaging and quantification of biofilms were performed using a crystal violet assay. All 92 MDR A. baumannii isolates involved in our study contained the bla_OXA-23 and bap genes. The average absorbance of biomass in Bap-producing strains was much greater than that in non-Bap-producing strains. In our study, only three REP-PCR types were found, and the isolates showing type A or type B were found more than 60 times among unique patients during the 5 years of surveillance. These results suggest that the isolates have persisted and colonized for 5 years, and biofilm formation ability has been responsible for their persistence and colonization.

• Korean Journal of Agricultural Science
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• v.45 no.4
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• pp.575-582
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• 2018

Bacterial fruit blotch (BFB) caused by Acidovorax citrulli is a devastating disease found in many cucurbits cultivation fields. The genetic diversity for 29 strains of A. citrulli collected from various cucurbits in South Korea was determined by DNA fingerprinting with a pathogenicity test, multi locus analysis, Rep-PCR (repetitive sequence polymerase chain reaction), and URP (universal rice primers) PCR bands. Two distinct groups (Korean Clonal Complex, KCC1 and KCC2) in the population were identified based on group specific genetic variation in the multi locus phylogeny using six conserved loci and showed a very high similarity with DNA sequences for representative foreign groups [the group I (CC1-1 type) and the group II (CC2-5 type)] widely distributed worldwide, respectively. Additionally, in the case of phaC, a new genotype was found within each Korean group. The KCC1 was more heterogeneous compared to the KCC2. The KCC1 recovered mainly from melons and watermelons (ratio of 6 : 3) and 15 of the 20 KCC2 strains recovered from watermelons were dominant in the pathogen population. Accordingly, this study found that two distinct groups of differentiated A. citrulli exist in South Korea, genetically very similar to representative foreign groups, with a new genotype in each group resulting in their genetic diversity.

• Korean Journal of Clinical Laboratory Science
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• v.47 no.2
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• pp.65-70
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• 2015

Antimicrobial agents or additives have commonly been used in domestic animals for the prevention and treatment of bacterial diseases. Unfortunately, this has resulted in the overgrowth of bacteria that is resistant to antimicrobial agents used by humans, and these might get disseminated to humans via the food. In this study, we investigated the prevalence of integrons, and characterized gene cassette arrays in Proteus mirabilis isolates obtained from chickens in Chungcheong province of Korea. Additionally, the correlation between gene cassette arrays and antimicrobial resistance rate was studied. A total of 26 Proteus mirabilis isolates were recovered from chickens in Chungcheong province in Korea. Antimicrobial susceptibility was determined by disk diffusion method. PCR and DNA sequencing were performed to characterize the gene cassette arrays. In addition, we employed repetitive extragenic palindromic sequence-based PCR (REP-PCR) method for clonality analysis of P. mirabilis strains. Of the 26 P. mirabilis isolates tested, 14 (53.8%) isolates carried class 1 integrons, while class 2 and class 3 integrons were not detected in our study. The class 1 integrons harbored genes encoding resistance to aminoglycosides (aacCA5, aadA2, aadA5 and aadA7), trimethoprim (dfrA17, and dfrA32), lincosamides (linF) and erythromycin (ereA). In particular, the presence of class 1 integron had a significant correlatation to a high resistance rate of aminoglycoside and trimethoprim. We confirmed that class 1 integrons are widely disseminated in P. mirabilis isolates from chickens, contributing to the resistance to diverse antimicrobial agents in Korea. To prevent further spreading of antimicrobial resistant genes among P. mirabilis isolates, constant monitoring and clinical policing will become necessary.

• The Plant Pathology Journal
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• v.24 no.4
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• pp.423-432
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• 2008

The difference of carbon source utilization and fatty acid composition between the pathotypes of Xanthomonas strains, which causing citrus bacterial canker was compared, and the physiochemical characteristics were used to analyze relationship of the strains for the first time. The pattern of several carbon sources utilization and fatty acids composition reliably discriminated the pathotypes of Xanthomonas strains. The dendrogram which was constructed by 95 carbon source utilization profiles differentiated X. axonopodis pv. citri A, $A^*$ and $A^w$ from the other pathotypes. When the dendrogram was drawn by combined analysis of carbon source utilization pattern and fatty acid composition, X. axonopodis pv. aurantifolii B, C and X. axonopodis pv. citrumelo formed a distinct cluster. The difference of carbon source utilization and fatty acid composition could be used effectively for the identification of pathotypes of citrus bacterial canker. The physiochemical characteristics strongly indicated that the strains isolated in Korea belong to X. axonopodis pv. citri A type. The cluster analysis by the band patterns of ERIC-, BOX- and REP-PCR allowed the discrimination of the pathotypes isolated from Korea. However, the rep-PCRs could not differentiate X. axonopodis pv. citri A types from $A^*$ and $A^w$ types. The overall results of metabolic profiles and rep-PCRs strongly indicated that the Korean isolates are X. axonopodis pv. citri A type.