• Journal of Microbiology and Biotechnology
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• v.33 no.3
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• pp.329-338
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• 2023

Enterohemorrhagic Escherichia coli (EHEC) is a foodborne pathogen that produces attaching and effacing lesions on the large intestine and causes hemorrhagic colitis. It is primarily transmitted through the consumption of contaminated meat or fresh produce. Similar to other bacterial pathogens, antibiotic resistance is of concern for EHEC. Furthermore, since the production of Shiga toxin by this pathogen is enhanced after antibiotic treatment, alternative agents that control EHEC are necessary. This study aimed to discover alternative treatments that target virulence factors and reduce EHEC toxicity. The locus of enterocyte effacement (LEE) is essential for EHEC attachment to host cells and virulence, and most of the LEE genes are positively regulated by the transcriptional regulator, Ler. GrlA protein, a transcriptional activator of ler, is thus a potential target for virulence inhibitors of EHEC. To identify the GrlA inhibitors, an in vivo high-throughput screening (HTS) system consisting of a GrlA-expressing plasmid and a reporter plasmid was constructed. Since the reporter luminescence gene was fused to the ler promoter, the bioluminescence would decrease if inhibitors affected the GrlA. By screening 8,201 compounds from the Korea Chemical Bank, we identified a novel GrlA inhibitor named Grlactin [3-[(2,4-dichlorophenoxy)methyl]-4-(3-methylbut-2-en-1-yl)-4,5-dihydro-1,2,4-oxadiazol-5-one], which suppresses the expression of LEE genes. Grlactin significantly diminished the adhesion of EHEC strain EDL933 to human epithelial cells without inhibiting bacterial growth. These findings suggest that the developed screening system was effective at identifying GrlA inhibitors, and Grlactin has potential for use as a novel anti-adhesion agent for EHEC while reducing the incidence of resistance.

• Korean Journal of Plant Tissue Culture
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• v.24 no.4
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• pp.225-231
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• 1997

For genetic engineering to be commercially viable, an efficient transformation system is needed to produce transgenic plane from diverse genotypes ("generalized protocol"). Development of such a system requires optimization of a number of components such as gene transfer agent, plant tissues competent for both regeneration and transformation, and control of transgene expression. Although several novel gene transfer methods have been developed for plane, a majority of stably transformed plane express the introduced genes at low levels. Moreover, silencing of selectable marker genes shortly after their incorporation into plant chromosomes may result in low recovery of transgenic tissues from selection. Matrix attachment regions (MARs) are DNA sequences that bind to the cell's proteinaceous nuclear matrix to form DNA loop domains. MARs have been shown to increase transgene expression in tobacco cells, and reduce position in mature transgenic plants. Flanking an antibiotic resistance transgene with MARs should therefore lead to improved rates of transformation in a diversity of species, and may permit recalcitrant species and genotypes to be successfully transformed. Literature review and recent data from my laboratory suggest that MARs can serve as a transformation booster in recalcitrant plant species.

• Journal of Veterinary Clinics
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• v.20 no.3
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• pp.302-307
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• 2003

Although, in human medicine, strains of methicillin-resistant staphylococi have become the most important causative agents of nosocomial infections, studies on the small animals are very. limited. The aim of this study was to determine mecA gene and susceptibility to antibiotics of staphylococci strains isolated from clinically ill or healthy dogs and cats, during the period August 2002-July 2003. A total of 136 staphylococci (87 coagulase-positive and 49 coagulase-negative) were investigated for antibiotic resistance, using disk diffusion and minimum inhibitory concentration (MIC) test. The mecA gene was detected using the polymerase chain reaction. The isolates belonged to the species S. aureus (53 isolates), S. intermedius (34 isolates), S. epidermidis (26 isolates) and other coagulase-negative staphylococci (CNS, 23 isolates). Of the 136 isolates, 43 (31.6%) were mecA-positive and the frequency of the ,presence of mecA gene varied among the different species. All S. aureus strains were mecA-negative and were found to be susceptible, with an oxacillin MIC $\leq$1 $\mu\textrm{g}$/ml. Five (13.6%) isolates of 36 that exhibited oxacillin resistance on the MIC testing were found to be mecA-negative, suggesting not all mecA-positive strains may be an oxacillin resistant. However, the mecA presence of the strains was correlated with high oxacillin resistance: 71.4% (10 isolates of 14; P < 0.001) for mecA-positive S. intermedius and 72.4% (21 isolates of 29; P < 0.001) for mecA-positive CNS isolates. About 69% (94 isolates of 136) showed resistance to at least one drug, and 22.8% (31 isolates) were resistant to four or more different drug classes. Resistance (36 isolates, 71.7%) to penicillin G was a common finidng. This study suggest that the mecA-positive staphylococci are prevalent in small animals, and selection of antibiotics to treat infections caused by mecA-positive staphylococci may be very limited because of multi-drug resistance.

• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.367-373
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• 2021

The antimicrobial resistance of Staphylococcus spp. and Gram negative strains present in air samples from waste sorting facilities was assessed. Phenotypic studies have revealed a high percentage of strains of Staphylococcus spp. resistant to methicillin. Genotypically and by RT-PCR, it was found that the mecA gene usually associated with methicillin resistance was present in 8% of the Staphylococcus strains isolated. About 30% of the Gram negative strains from the same samples also displayed resistance to meropenem and 79% of these were resistant to multiple antibiotics from different classes, namely cephalosporins and β-lactams. The results suggest that in professional activities with high levels of exposure to biological agents, the quantification and identification of the microbial flora in the work environment, with the determination of the presence of potential agents displaying multi-resistances is of relevance to the risk assessment. The personal protection of workers is particularly important relevance in these cases, since many of the strains that exhibit multi-resistance are potential opportunistic agents.

• Korean Journal of Microbiology
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• v.36 no.3
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• pp.216-220
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• 2000

Eighty-eight strain3 o~methicillin resistant Stopllylococcus awecis were Isolated from pus (64.7%); spuhm (26.2%), blood, fluid, andurine of 83 patients at Dong-A Hospital in P~~san to invesligate theil-coagulase typ- Ing, and multi-drug resistaut ppattems. The presence of niec A gene confe~~ing melhicillin resistance was tested by polymerase chain reaction (PCR) with uwo mec A gene specific primers using purified clromosonlal DNA as templates. DNA fragments of expected size wel-e detected frorn 86 strains, but not from two strains. !i coagulase typmg, the 86 isolates were assigned to 5 coagulase lypes, I, 11, lll. 1V, VI, VII, VIlI, but there was no isolate helong lo type V. The most abundant coagulase type was type TI(50 %), lollowed by type IV Rest ofthe coagulase types were ininor; ranging fmm 4.5 to 12.5 '% Most of the type I1 ~netlucillin resistant Stapl\ulcorneryiococcus nwem (MRSA) strams were isolated from the generd sulzely ward, but major strains of type IV were Isolated from the otorhinolq~ngology of the hospital's outpatient clinic center. All of the 88 st~nins were sensitive to vancomycin and teicoplanin, but 71 (81%) strains showed multi-drug resitant to penicillin, cephalotl~n, eiythroinycin, gentan~ycin, imipenem, clindamycin, ciprofloxacin and ooxacillin. Yo relationship was found between the antibiotic resistance pattems aud the coagulase typing patterns.

• Research in Plant Disease
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• v.28 no.1
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• pp.26-31
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• 2022

Pseudomonas syringae pv. actinidiae (Psa) is the causal agent responsible for the bacterial canker disease of kiwifruit plants. Psa strains are divided into five different biovars based on genetic and biochemical characteristics. Among them, biovar 2 and 3 strains of Psa were isolated and have been causing widespread damages in Korea. One of the most effective ways to control Psa is to use an antibiotic such as streptomycin. However, Psa strains resistant to this antibiotic were isolated in Korea, and an earlier study revealed that the resistance in the biovar 2 is associated with strA-strB genes. This study aimed to determine the molecular resistance mechanism of Psa biovar 3 strains to streptomycin. Sequencing the rpsL gene encoding ribosomal protein S12 from three streptomycin-resistant strains screened in the laboratory revealed that a spontaneous mutation occurred either at codon 43 or 88. Meanwhile, in four streptomycin-resistant strains of Psa biovar 3 isolated from two kiwifruit orchards, a single nucleotide in codon 43 of the rpsL, which is AAA in streptomycin-sensitive strain, was substituted for AGA causing an amino acid change from lysine to arginine. The resistant mechanism in all biovar 3 strains obtained in Korea was identified as a mutation of the rpsL gene.

• Journal of Life Science
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• v.21 no.2
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• pp.257-264
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• 2011

Seventy five methicillin- resistant Staphylococcus aureus (MRSA) strains and 24 methicillin- susceptible S. aureus (MSSA) were isolated from clinical specimens obtained from a hospital in Suncheon, Jeonnam province, Korea, from July to December, 2009. Antibiotic resistance was determined using the disc diffusion method. Genes encoding enterotoxin (SE), toxic shock syndrome toxin-1 (TSST-1), exfoliative toxin (ET) and Panton-Valentine leukocidin (PVL) were detected by multiplex PCR-mediated amplification using specific primers. Sixty (80%) MRSA isolates possessed either one or more toxin genes and the most common pattern that coexisted in MRSA was seb, sec, seg, sei and tst (22.7%) followed by coexistence of sec, seg, sei and tst genes (18.7%). Gene pvl encoding leukocidin was not found. Significant correlation between the production of sec, seg, sei and tst genes was found. MRSAs were resistant to erythromycin (89% of the isolates), gentamicin (70.7%), ciprofloxacin (69.3%), clindamycin (61.3%) and tetracycline (58.7%), while MSSAs were susceptible to the antibiotics with the exception of erythromycin. Toxin genes seb, sec and tst were related to the tetracycline resistance of MRSA.

• Journal of Food Hygiene and Safety
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• v.39 no.3
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• pp.266-272
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• 2024

This study assessed the contamination level of food poisoning bacteria on handles of food service equipment in childcare centers to prevent food poisoning and analyzed toxin genes and antibiotic resistance of isolated strains. The isolates used in this study were collected from 101 childcare centers in Jeollanam-do. Four strains of Bacillus cereus and two strains of Staphylococcus aureus were isolated on the handles of food service equipment (refrigerators and freezers). The toxin genes of B. cereus were detected as nheA, nheB, nheC, entFM, and cytK. No toxin genes of S. aureus were detected. B. cereus showed resistance to β-lactam antibiotics, such as ampicillin and cefepime. S. aureus also showed resistance to antibiotics such as ampicillin and cefepime. Therefore, microbial safety and hygiene management, such as periodic sterilization of handles, should be strengthened to prevent food poisoning caused by cross-contamination of food service equipment handles in childcare centers.

• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1460-1466
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• 2015

In this study, the prevalence of Shiga toxin-producing Escherichia coli (STEC) was investigated among raw meat or meat products from slaughterhouses and retail markets in South Korea, and their potential for antibiotic resistance and virulence was further analyzed. A total of 912 raw meats, including beef, pork, and chicken, were collected from 2008 to 2009. E. coli strains were frequently isolated in chicken meats (176/233, 75.9%), beef (102/217, 42.3%), and pork (109/235, 39.2%). Putative STEC isolates were further categorized, based on the presence or absence of the Shiga toxin (stx) genes, followed by standard O-serotyping. Polymerase chain reaction assays were used to detect the previously defined virulence genes in STEC, including Shiga toxins 1 and Shiga toxin 2 (stx1 and 2), enterohemolysin (ehxA), intimin (eaeA), STEC autoagglutination adhesion (saa), and subtilase cytotoxin (subAB). All carried both stx1 and eae genes, but none of them had the stx2, saa, or subAB genes. Six (50.0%) STEC isolates possessed the ehxA gene, which is known to be encoded by the 60-megadalton virulence plasmid. Our antibiogram profiling demonstrated that some STEC strains, particularly pork and chicken isolates, displayed a multiple drug-resistance phenotype. RPLA analysis revealed that all the stx1-positive STEC isolates produced Stx1 only at the undetectable level. Altogether, these results imply that the locus of enterocyte and effacement (LEE)-positive strains STEC are predominant among raw meats or meat products from slaughterhouses or retail markets in Korea.

• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.2100-2105
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• 2015

Probiotic bacteria must have not only tolerance against bile salt but also no genes for antibiotic resistance. Leuconostoc citreum is a dominant lactic acid bacterium in various fermented foods, but it is not regarded as a probiotic because it lacks bile salt resistance. Therefore, we aimed to construct a bile salt-resistant L. citreum strain by transforming it with a bile salt hydrolase gene (bsh). We obtained the 1,001 bp bsh gene from the chromosomal DNA of Lactobacillus plantarum and subcloned it into the pCB4170 vector under a constitutive P710 promoter. The resulting vector, pCB4170BSH was transformed into L. citreum CB2567 by electroporation, and bile salt-resistant transformants were selected. Upon incubation with glycodeoxycholic acid sodium salt (GDCA), the L. citreum transformants grew and formed colonies, successfully transcribed the bsh gene, and expressed the BSH enzyme. The recombinant strain grew in up to 0.3% (w/v) GDCA, conditions unsuitable for the host strain. In in vitro digestion conditions of 10 mM bile salt, the transformant was over 67.6% viable, whereas only 0.8% of the host strain survived.