• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.5
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• pp.796-801
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• 2001

A total of 277 mineral spring water samples in Kangwon province from 1999 to 2000 were analyzed for the presence of Yersinia spp. by the conventional Food and Drug Administration protocol, and presumptive strains were identified by morphological, cultural and biochemical tests according to Bergey’s manual. Also, the biotypes, serotypes, and susceptibility to 12 antibiotics were tested. Among the total 277 mineral spring water samples, 40 samples (14.4%) were found to be contaminated with Yersinia species. Among the 40 strains of Yersinia spp. isolates, 33 strains (82.5%) for Yersinia enterocolitica, 4 strains (10%) for Yersinia frederiksenii, 2 strains (5%) for Yersinia intermedia, and 1 strain (2.5%) for Yersinia sakazaki were identified, respectively. Of 40 Yersinia spp. isolates, Yersinia enterocolitica (82.5%) was the most predominant species in the mineral spring water samples compared to other Yersinia species. Compared to direct culture method after KOH treatment and KOH treatment method after cold enrichment for better isolation ratio of according to comparision of Yersinia species, the detection ration (18.5%) of KOH treatment method after cold enrichment was about 3 times better than that (6.1%) of direct culture method after KOH treatment. According to serotypes of Y. enterocolitica isolates, O : 5 (12.9%) was the most predominant and followed by O : 3 (9.7%), O : 8 (6.5%), and O : 9 (3.2%), and others. For biotypes of Y. enterocolitica isolates, 1A (71.0%) was the most predominantly abundant and followed by 3A (12.9%), 3B (9.7%), 1B (3.2%) and 5 (3.2%). Also, an antibiotic susceptibility test showed that Yersinia spp. isolates were very susceptible to the antibiotics tested, but they were very strongly resistant to ampicillin, cephalothin and carbenicillin.

• Korean Journal of Medicinal Crop Science
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• v.15 no.4
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• pp.285-290
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• 2007

This study was conducted to introduce phosphinothricin acetyl transferase (PAT) gene, resistant to basta which was non-selective herbicide, into balloon flower (Platycodon grandiflorum A. De. candolle). Seeds were germinated on MS medium, and 10-day-old immature cotyledon explants and 30-day-old leaf explants were cocultured with Agrobacterium tumefaciens strain MP 90 (pBinSyn) on 1/10 MS medium for 48 hours in the dark at $25^{\circ}C$. The cultures were transferred for selection of kanamycin-resistant shoots to the MS medium supplemented with 0.2 $mg/{\ell}$ NAA, 1.0 $mg/{\ell}$ BA, 3% sucrose, 100 $mg/{\ell}$ kanamycin, 500 $mg/{\ell}$ carbenicillin. Shoots were obtained from 10-day-old immature cotyledon explants after 4 weeks of culture. The shoots were subcultured twice every 4 weeks on the same medium for growth of transgenic shoots. Successful transformation was confirmed by histochemical GUS assay, PCR analysis, RT-PCR analysis, 10 $mg/{\ell}$ phosphinothricin treatment and 0.3% basta spray. The basta-resistant transgenic plants flowered normally.

• Journal of Life Science
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• v.10 no.3
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• pp.262-272
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• 2000

As a part of investigation for basic epidemiology of diarrheogenic disease, we attempted isolation of Escherichia coli from patients with diarrhea. Seven hundred and twenty-one strains of E. coli were isolated from 1,239 patients with diarrhea. Seasonal distribution of patient with diarrhea was shown the most high at August (18.2%). Age group distribution of patient was shown the most high at children (54.6%, 2 to 10 years old). The serotypes of 721 E. coli isolates were in order of serotype O44 (16.8%), O153 (8.6%), O1 (7.5%), O166(5.7%), O8 and O86a (4.7%), and O125 (4.6%). The supernates cultured 36 strains among 721 E. coli isolates were indicated cytotoxicity against monolayered Vero cells. All of the isolates were susceptible to amikacin. The isolates were resistant in order of novobiocin (99.0%), moxalactam (97.1%), carbenicillin (96.1%), tetracycline (90.4%), ampicillin (85.9%), gentamicin (84.0%), streptomycin (78.4%), cephalothin (46.6%) and polymyxin B (4.2%). In the antibiotic resistant patterns, 125 kinds of multiple resistance patterns of E. coli isolates were detected. The highest resistant pattern was ampicillin-carbenicillin-chloramphenicol-cephalothin-erythro-mycin- gentamicin-moxalactam-novobiocin-penicillin G-streptomycin-tobramycin-tetracycline-tri methoprim type (24.3%).

• Journal of Plant Biotechnology
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• v.33 no.1
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• pp.27-32
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• 2006

To clarify the roles of bromelain in plants, we isolated BL1 gene encoding bromelain from pineapple stem tissues and sequenced. The full length cDNA is 933 bp and encodes a polypeptide of 311 amino acid residues. The cDNA is most similar 94% at the amino acid level to bromelain previously isolated from pineapple (BAA21929). Explants of Lactuca sativa were co-cultivated with Agrobacterium tume-faciences LBA 4404 strains containing nptII and BL1 gene for transformation. Through initial selection of regenerated explants by culturing on a kanamycin and carbenicillin containing MS medium, multiple shoots were obtained after 2 months of culture. For a complementary step of selection, putative transgenic shoots were transferred to 1/2 Ms basal medium supplemented with 100 mg/L kanamycin and 500 mg/L carbenicillin. The selected shoots were obtained T1 generation seeds with emasculation, and tested with PCR analysis using 35S promoter and BL1 specific primers whether BL1 gene was introduced to genome of the plants. These results confirmed that produced the specific PCR bands in the putative transgenic lines. Additionally the Northern blot and endo protease activity showed that transcripts of BL1 gene were detected in transgenic lines. Theses results suggest that BL1 gene be successfully integrated and transcripted in the transgenic lettuce plants.

• Journal of Food Hygiene and Safety
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• v.15 no.3
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• pp.241-247
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• 2000

The incidence of verotoxin-producing Escherichia coli(VTEC) was surveyed in domestic foods including hamburger, raw meats and vegetables from 1997 to 1999. The molecular biological characteristics of the isolates were analyzed. Three VTEC strain were isolated from 1,700 samples. Serotypes of those isolates were 0157 : H7, 026 H4, and 056 : Hl2, respectively. Serotype O26 : H4 produced VT I and VT II, and 055 Hl2 isolate produced VT I, however the 60 MDa plasmid DNA and eae gene were not found from both strains. One 0157 : H7 isolate produced VT II and harbour 60 MDa plasmid DNA, however eae gene was not found in the strain. Although they produced VT, it seemed that the virulence of two strains were relatively weak because of the lack of the eae gene. In addition, the serotype O157 : H7 isolate resistant to ampicillin and streptomycin, while isolates of serotype O26 : H4 and O55 : Hl2 were multi-resistant to antibiotics including ampicillin, carbenicillin , cephalothin, trimethoprim/sulfamethoxazole and tetracycline. Supernatants of cultures of all three isolates were showed cytotoxic effect to vero and HeLa cell

• Korean Journal of Food Science and Technology
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• v.31 no.1
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• pp.183-188
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• 1999

The incidence of Yersinia enterocolitica in raw meat was determined over 10 month period. Y. enterocolitica was isolated from 8.5% of beef, 17.0% of pork and 25.6% of chicken. Overall prevalence of Y. enterocolitica in raw meat was 17.5%. Seasonal difference was observed in isolation rate in which pork and chicken samples collected in the second half of the year twice was more than that of the first half of the year. Serotypes of Y. enterocolitica isolates were O:5 (17.3%), O:8 (8.6%), O:3 (6.2%), O:1,2 (1.2%), and others. The antibiotics susceptibility tests showed Y. enterocolitica isolates were resistant to carbenicillin, ampicillin, erythromycin, penicillin and bacitracin. In contrast it showed sensitivity to polymyxin B, kanamycin, ciprofloxacin, gentamicin, trimethoprim/sulfamethoxazole, nalidixic acid, and tetracycline. PCR with specific primers derived from ail gene of Y. enterocolitica was applied to detect and confirm pathogenic Y. enterocolitica. About 10% of the isolated Y. enterocolitica proved to be pathogenic and most of them were found in pork. However, proper cooking and meat process can kill and remove all Y. enterocolitica in meat, because the organism is very sensitive to heat.

• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.95-99
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• 1995

We have previously established a system for plant regeneration through somatic embryogenesis and Agrobacterium-mediated transformation of Korean ginseng. In this study to produce a fungus-resistant plant, we introduced a bean chitinase gene into ginseng using the transformation system. A binary vector pChi/748 was constructed by introducing the bean basic chitinase gene into EcoRI site of pGA748 which carries the CaMV 35S promoter governing the introduced gene and neomycin phosphotransferase II(NPT-II)gene as a positive selection marker. Cotyledonary explants were cocultured with A. tumefaciens strain LBA4404 harboring the binary vertor pChi/748 for 48 h, and transferred to MS medium supplemented with l mg/L2,4-D,0.1mg/L kinetin, 100 mg/L kanamycin, and 500mg/L carbenicillin. Kanamycin-resistant calli were formed on the cut surface of cotyledonary explants after one month of culture, and subsequently they gave rise to somatic embryos. Upon transfer onto medium containing 1 mg/L each of BA and GA$_3$, most of them converted to plantlets after 5 weeks of culture. The genomic DNA of eight kanamycin-resistant regenerants was subjected to polymerase chain reaction (PCR) using two specific 21-mer oligonucleotides derived from the chitinase gene. PCR-Southern blot analysis confirmed that the chitinase gene was incorporated into six out of the eight regenerants..

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.93-98
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• 2002

A VP6 fragments was subcloned with BamHI in the binary pMBP-1 vector under Califlower Mosaic Virus (CaMV) 355 promoter and neomycin phosphotransferase II (npt II) gene. The recombinant binary vector was mobilized into Agrobacterium-tumefaciens LBA4404 by the freeze-thaw method and potato (Solanum tubensum L. cv Desiree) was transformed by modified leaf-disc cocultivation. Shoots were induced on MS medium with 0.01 mg/L NAA, 0.1 mg/L GA$_3$, 2.0 mg/L Zeatin, 100.0 mg/L kanamycin, 500.0 mg/L carbenicillin. In order to identify the copy number of VP6 into potato plant, total genomic DNA was isolated from transgenic potato and analysed by Southern blotting. Genomic DNA and total mRNA analysis demonstrated the incorporation of the foreign gene into the potato genome, as well as their transcription.

• Korean Journal of Veterinary Research
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• v.40 no.4
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• pp.763-767
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• 2000

This study was performed to isolate and identify bacteria from uterus with pyometra and examine their susceptibility to antimicrobial agents. Uterus of 16 bitches with pyometra were surgically removed by ovariohysteroctomy and then bacteria were isolated and identified. Also, susceptibility test to 15 antimicrobial agents was performed. Out of 16 bitches, 11 strains of Escherichia coli, 2 strains of Serratia marcescens, and 1 strain of Staphylococcus aureus and Salmonella spp. were identified. In antimicrobial susceptibility test, the majority of isolates were susceptible to enrofloxacin, norfloxacin, chloramphenicol, nalidixic acid, gentamicin, trimethprim-sulfamethazole, tetracycline, and moderately susceptible to carbenicillin, amikacin, ampicillin, neomycin, but resistant to vancomycin, streptomycin, bacitracin and colistin. In conclusion, E coli was the most common bacteria isolated from bitches with pyometra and those susceptible antimicrobial agents could be recommended to medical therapy of pyometra.

• Journal of Korean Society of Forest Science
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• v.79 no.3
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• pp.278-284
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• 1990

We have demonstrated expression of bacterial genes transferred into cells of Populus nigra ${\times}$ P. maximowiczii by A. tumefaciens strain 6044 (pGA 472). We determined the optimum concentration of kanamycin sulfate for effective selection of punctured leaf transformed using Agrobacterium binary vector pGA 472 containing a neomycine phosphotransferase gene (NPT-II) which confers kanamycin resistance. The combination of cefotaxime (200mg/l) and carbenicillin (300mg/l) showed good performance of discarding Agrobacterium from inoculated punctured leaf. A relatively low concentration (10mg/l) of kanamycin sulfate inhibited callus and shoots induction from punctured leaf. Number of shoots regenerated from co-cultured punctured leaf was 3.0 on MS basal medium supplemented with 10 mg/l kanamycin sulfate, while that of not co-cultured punctured leaf was none. The regeneration rate was 10% from the punctured leaf co-cultured on MS medium with 10 mg/l kanamycin. Regenerated shoots are developing from micropropagation for Southern blot analysis and inheritance of the kanamycin resistance trait (NPT-II).