• Korean Journal of Food Science and Technology
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• v.53 no.6
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• pp.799-808
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• 2021

This study was performed to investigate the extent of microbial contamination, the presence of enterotoxin genes, and the antibiotic susceptibility of Bacillus cereus in 58 red pepper plants and 43 environmental samples (soil, irrigation water, and gloves) associated with the plant cultivation. The detected counts of total aerobic bacteria, coliform bacteria, Escherichia coli, Bacillus cereus, and Staphylococcus aureus were lower in these samples, as compared to the regulations of standards for foods; moreover, pathogens, such as E. coli, E. coli O157:H7, Listeria monocytogenes, and Salmonella spp., were not detected. Genes encoding hemolysin BL enterotoxins (hblA, hblC, and hblD) as well as non-hemolytic enterotoxins (nheA, nheB, and nheC) were detected in 23 B. cereus specimens that were isolated from the test samples and had β-hemolytic activity. Interestingly, B. cereus is resistant to β-lactam and susceptible to non-β-lactam antibiotics. However, in this case, the isolated B. cereus specimens exhibited a shift from resistant to intermediate in response to cefotaxime and from susceptible to intermediate in case of rifampin, trimethoprim-sulfamethoxazole, vancomycin, clindamycin, and erythromycin. Therefore, the levels of B. cereus should be monitored to detect changes in antibiotic susceptibility and guarantee their safety.

• The Journal of Natural Sciences
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• v.4
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• pp.103-142
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• 1991

These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

• Pediatric Infection and Vaccine
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• v.14 no.1
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• pp.83-90
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• 2007

Purpose : Staphylococcal scalded skin syndrome (4S) is a well known disease defined by clinical, microbiological and histological criteria caused by Staphylococcus aureus. This disease is uncommon but has been increasingly recognized. We investigated the clinical features of staphylococcal scalded skin syndrome. Methods : We reviewed retrospectively medical records of 53 patients diagnosis of staphylococcal scalded skin syndrome who were admitted to Changwon Fatima hospital from February 2002 to December 2005. These patients were divided into 3 clinical types; generalized type, intermediate type, abortive type. Age, sex ratio, clinical manifestations, laboratory findings, response to therapy and prognosis were investigated. Result : 1)The mean age of patients was 2.8 years, ranging from 20 days to 7 years. Male-to-female ratio was 1.9:1. 2) By clinical types, 6 patients were in the generalized type (11%), 29 patients in the intermediate type (55%), 18 patients in the abortive type (34%). The coexisting diseases were variable, including conjunctivitis (25 cases), atopic dermatitis (11 cases), otitis media (1 case). On laboratory findings, most of patients didn't have leukocytosis or increased C-reactive protein. 4) A total of fifteen Methicillin Resistant Staphylococcal Aureus (MRSA) strains were isolated from September 2003 through December 2005. Fourteen strains were positive for exfoliative toxin B gene by PCR and negative for enterotoxin, toxic shock syndrome toxin and Panton-Valentine leukocidin genes. 5) The mean duration of admission was 7.3 days. Patients were treated with vancomycin or amoxacillin/clavulanate or ampicillin/sulbactam or cefuroxime without significant sequelaes. Conclusion : Recently, Staphylococcal scalded skin syndrome caused by exfoliative toxin B produced by MRSA in the Changwon area has been increasing.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.65 no.3
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• pp.172-181
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• 2020

Koshihikari has been one of the most popular rice cultivars with good eating quality since the 1960s despite its susceptibility to blast disease and lodging. To map the genes controlling blast resistance and to develop promising blast-resistant breeding lines inheriting Koshihikari's high eating quality, a recombinant inbred line (RIL) population was developed from a cross between Koshihikari and a blast resistance donor with early maturity, Baegilmi. A total of 394 Koshihikari × Baegilmi RILs (KBRIL), and the two parents, were evaluated for blast resistance and major agronomic traits including heading date, culm length, panicle length, and tiller number. A linkage map encompassing 1,272.7 cM was constructed from a subset of the KBRIL (n = 142) using 130 single nucleotide polymorphisms. Two quantitative trait loci (QTL) for blast resistance, qBL1.1 harboring Pish/Pi35 and qBL2.1 harboring Pib, were mapped onto chromosomes 1 and 2, respectively. qBL1.1 was detected in both of the experimental sites, Namwon and Jeonju, while qBL2.1 was only detected in Namwon. qBL1.1 and qBL2.1 did not affect agronomic traits, including heading date, culm length, panicle length, and tiller number. From the 394 KBRILs, lines that were phenotypically similar to Koshihikari were selected according to heading date and culm length and were further divided into the following two groups based on blast resistance: Koshishikari-type blast resistant lines (KR, n = 15) and Koshishikari-type blast susceptible lines (KS, n = 15). Although no significant differences were observed in the major agronomic traits between the two groups, the KR group produced a greater mean head rice ratio than the KS group. The present study provides useful materials for developing blast-resistant cultivars that inherit both Koshihikari's high eating quality and Baegilmi's blast resistance.

• Korean Journal of Microbiology
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• v.49 no.3
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• pp.228-236
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• 2013

Campylobacter jejuni is one of important food-borne pathogens causing human gastroenteritis. We isolated 42 strains of C. jejuni from diarrhea patients and 4 food-poisoning outbreaks in 2010, Gyeonggi-do. In this study, 42 strains were tested for genetic characteristics, the serotype distribution and antimicrobial resistant rate. The presence of hipO (100%), cdtB (100%), and mutated gyrA (95.2%) genes was detected in C. jejuni by polymerase chain reaction (PCR). Detection of mutated gyrA gene correlated with ciprofloxacin resistance. Forty isolates had mutated gyrA gene and were actually resistant to ciprofloxacin. Furthermore, comparing the gyrA DNA sequence data, ciprofloxacin-resistant isolates had a mutation of the DNA sequence from ACA (threonine) to ATA (isoleucine). But 41 strains (97.6%) of patient isolates were susceptible to erythromycin and azithromycin. A total of 35.7% among 42 C. jejuni isolates were identified into 4 different serotypes. The serotype distribution of C. jejuni strains were shown to be HS2(B), HS3(C), HS4(D), HS19(O). To investigate the genotypes of C. jejuni isolated in Gyeonggi province, repetitive sequence polymerase chain reaction (rep-PCR) analysis and SmaI-digested pulsed-filed gel electrophoresis (PFGE) profile analysis were performed. From the PFGE analysis of 42 C. jejuni strains, 12 clusters of PFGE profile were obtained. On the other hand, 11 clusters of rep-PCR profile were obtained from 42 strains of C. jejuni.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.401-406
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• 2017

In this study, using the VITEK 2 system, 74 samples (22.6%) out of 327 specimens were identified by the growth of Enterococcosel media (EV6 agar) supplemented with $6{\mu}g/mL$ of vancomycin. Enterococcus faecium was identified as 55 strains (74.3%), Enterococcus casseliflavus as 2 strains (2.7%), Enterococcus avium as 1 strain (1.4%), and Enterococcus gallinarum as 16 strains (21.6%). Among the 55 phenotypes of Enterococcus faecium, 42 (76.4%), 9 (16.4%), and 4 strains (7.3%) showed the vanA, vanB, and vanC phenotype, respectively. The 16 strains of Enterococcus gallinarum and 2 strains of Enterococcus casseliflavus showed the vanC phenotype and the 1 strain of Enterococcus avium had the vanB phenotype. The one strain of Enterococcus faecium propagated only in EV4 and was susceptible to both vancomycin and teicoplanin according to the antimicrobial susceptibility test using the VITEK 2 system. The vancomycin resistance phenotype gene was not detected by PCR. A total of 327 specimens were cultured in Enterococcosel broth supplemented with $6{\mu}g/mL$ of vancomycin (EV6 broth), and 120 strains (36.7%) were isolated. These 120 strains were subjected to vancomycin resistant genotyping by a multiplex real-time polymerase chain reaction and 51 strains (42.5%) showed vanA; 5 strains (4.2%) showed vanA and vanC; and 18 strains (15%) showed vanC. Vancomycin resistance genotypes were not detected in the remaining 46 strains (38.3%).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.2
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• pp.115-121
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• 2002

To develop a new method of conjugation and to determine the distribution of R plasimds, we isolated multi-drug resistant strains from fish pathogenic bacteria in the farms of south and east seacoasts of Korea. Out of the 134 isolates examined, 10 showed resistance to chloramphenicol, tetracycline, streptomycin, ampicillin, colistin, nalidixic acid, oxolinic acid and kanamycin. One out of 10 multi-drug resistance bacteria, Vibfio damsela JE1 (V. damsela JE1), contained transferable R plasmid of chlorarnphenicol- tetracycline resistance genes and other nucleic acids encoding ampicillin and kanamycin resistance. The presence of the R plasmid was confirmed by conjugation using the chromocult medium (CC) as a selective and differential medium for transconiugants with identification based on the growth or colors of the colonies. The frequency of R plasmid transfer with filter mating method was come out much higher than that of broth mating method and appeared to be dependent upon the mating time and temperature. The optimum conditions for filter mating method were found to be 30$^{\circ}C$ and 24hrs as mating temperature and period, respectively, Moreover, donor cells with R plasmid, both isolate and standard bacteria, were shown to have an ability to transfer the plasmid against Escherichia coli K-12 HB101 (E. coli HB101) and Edwardsiella tarda (E. tarda) RE14 at fairly high frequencies, finally, we isolated 3 isolates of Sphingomonas sp., carrying R plasmid from 12 multi-drug resistant bacteria in normal microflora of the flounder (Paralichthys olivaceus) group used for the isolation of V emsela JE1 four months before. The same size and gene transfer chayateristics of R plasimds with those of V damsela JE1 confirmed that normal microflora have the reservoir activity for R plasmid in natural aquatic environment.

• Tuberculosis and Respiratory Diseases
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• v.63 no.2
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• pp.128-138
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• 2007

Backgrounds: Mutations of katG and inhA (ORF and promoter) are known to be related to isoniazid (INH) resistance of Mycobacterium tuberculosis. Because reports on these mutations in Korean isolates are limited (i.e. only the frequency of katG codon 463 was evaluated.), we tried to know the kinds of mutations of two genes and their frequencies in INH resistant Korean M. tuberculosis strains. Methods: PCR was performed to amplify katG (2,223 bp), inhA ORF (-77~897, 975 bp), and inhA promoter (-168~80, 248 bp) from 29 multidrug resistant M. tuberculosis (MDR-TB) DNAs prepared by bead beater-phenol method. Their sequences were determined and analyzed by ABI PRISM 3730 XL Analyzer and MegAlign package program, respectively. Results: All of the isolates had more than one mutation in katG or inhA gene. Twenty seven (93%) of 29 tested strains had katG mutations, which suggests that katG is a critical gene determining INH resistance of M. tuberculosis. Amino acid substitutions, such as Arg463Leu and Ser315Thr, due to point mutations of the katG were the most frequent (62.1% and 55.2%) mutations. In addition, deletion of the katG gene was frequently observed (17.2%). Analyzed Korean MDR-TB isolates also had variable inhA mutations. Point mutation of inhA promoter region, such as -15 ($C{\rightarrow}T$) was frequently found. Substitution of amino acid (Lsy8Asn) due to point mutation ($AAA{\rightarrow}AAC$) of inhA ORF was found in 1 isolate. Interestingly, 14 point mutated types that were not previously reported were newly found. While four types resulted in amino acid change, the others were silent mutations. Conclusions: Although it is not clear that the relationship of these newly found mutations with INH resistance, they show marked diversity in Korean MDR-TB strains. It also suggests their feasibility as a molecular target to supplement determining the INH resistance of clinical isolates because of the possible existence of low-level INH resistant strains.

• Research in Plant Disease
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• v.28 no.2
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• pp.82-91
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• 2022

Recently, several in vitro studies have reported antiviral activity of agents of systemic acquired resistance against plant virus infection, but the approach has not been applied in a wide range of agricultural fields. The objective of this study was to evaluate the inhibitory effect of the exogenous application of salicylic acid (SA), chitosan (CH), or eugenol (EG) in tomato yellow leaf curl virus (TYLCV) infection of greenhouse-grown tomato plants. In vitro, the initial time of symptom was observed in TYLCV-infected plants (VP) of the resistant cultivar 'Superdotaerang' at 12 days post inoculation (dpi) after application of antiviral agents. At 32 dpi, the disease rate of TYLCV in the CHT+VP (0.1% chitosan and virus infected control) treated plants was 87.5%, lower than that of the other treatment. However, the virus content in the CHT+VP treated plants was higher than those of the other treatments, and SA, EG, and CH did not show significant effect on plant height or shoot and root fresh weight. Our results from summer-cultivated greenhouse-grown tomatoes show that none of the tested agents had an inhibitory activity on viral infection or yield of tomato 'Dotaerangsola'cultivar. In contrast, all treated 'TY Giants' cultivars that possessed TYLCV resistance genes Ty-1 and Ty-3a did not show typical symptoms and the virus content was remarkably lower than those in the TYLCV treated plants in 'Superdotaerang'. The results of this research indicated that the planting of resistant tomato cultivars was effective method instead of using SA, EG, and CH (known as resistance-inducing factors for control) of TYLCV in the field.

• Journal of Plant Biotechnology
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• v.41 no.2
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• pp.81-88
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• 2014

Salt cress (Thellungiella halophila or Thellungiella parvula), species closely related to Arabidopsis thaliana, represents an extremophile adapted to harsh saline environments. To isolate salt-tolerance genes from this species, we constructed a cDNA library from roots and leaves of salt cress plants treated with 200 mM NaCl. This cDNA library was subsequently shuttled into the destination binary vector [driven by the cauliflower mosaic virus (CaMV) 35S promoter] designed for plant transformation and expression via recombination- assisted cloning. In total, 305,400 pools of transgenic BASTA-resistant lines were generated in Arabidopsis using either T. halophila or T. parvula cDNA libraries. These were used for functional screening of genes involved in salt tolerance. Among these pools, 168,500 pools were used for primary screening to date from which 7,157 lines showed apparent salt tolerant-phenotypes in the initial screen. A secondary screen has now identified 165 salt tolerant transgenic lines using 1,551 (10.6%) lines that emerged in the first screen. The prevalent phenotype in these lines includes accelerated seed germination often accompanied by faster root growth compared to WT Arabidopsis under salt stress condition. In addition, other lines showed non-typical development of stems and flowers compared to WT Arabidopsis. Based on the close relationship of the tolerant species to the target species we suggest this approach as an appropriate method for the large-scale identification of salt tolerance genes from salt cress.