• Korean Journal of Microbiology
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• v.28 no.3
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• pp.236-242
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• 1990

To develop the new strains of microorganisms having the degradative ability for various aromatic hydrocarbons, the hybrid plasmid pKG2 having the 2,4-Dichlorophenoxyacetic acid(2,4-D) degradative genes, the hybrid plasmid pKG3 containg the naphthalene degradative genes and TOL plasmid were introduced into Pseudomonas putida KUD 12 and P. putida KUP 10 by transformation or conjugation which originally have the degradative ability of the synthetic surfactants and phthalate esters, respectively. From P. putida KUD12, the new strains of P. putida KUD101(pKG2), KUD102(pKG3), KUD103(TOL), and KUD202(pKG3, TOL) were obtained, and KUD106(pKG2), KUD107(pKG3), KUD108(TOL) were originated from the P.putida KUP10. The degradative abilities in P. putida KUD101, KUD102 and KUD107 were similar with those of the original strains. The P. putida KUD103, KUD106 and KUD202 had a little lower and P. putida KUD108 had a better degradative abilitie than those of the original ones. In the case of mixed cultures, the mixed culture of KUD107 and KUD108 had a better degradative abilities than those of the other mixed cultures.

• Korean Journal of Microbiology
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• v.25 no.4
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• pp.282-289
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• 1987

The characteristics of the plasmid pKU10 isolated from Pseudomonas putida KU816 were investigated and its restriction map was constructed. The pKU10 plasmid was a small R plasmid carrying genes for resistance to ampicillin, tetracyclin, and chloramphenicol, and cured by treatment with mitomycin C. The molecular size of pKU10 was estimated to be 9.4Kb. Pseudomonas strains and E. coli cells could be transformed for antibiotic resistance characters specified by pKU10 plasmid DNA. By incompatibility test with other plasmids, pKU10 is grouped into IncP-1. EcoRI, XhoI, SalI, BglII, and SmaI cleaved pKU10 once, while PstI cleaved at two sites, and HindIII cleaved at six sites. The restriction map was constructed by partial and complete digestion of the purified plasmid DNA with single, double, or triple restriction enzymes. Thus, pKU10 is expected to be used for a cloning vector in Pseudomonas cells.

• Korean Journal of Microbiology
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• v.34 no.4
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• pp.231-235
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• 1998

We have introduced a genetic assay to study the existence of intramolecular triplexes in Escherichia coli. A plasmid containing the gene that encodes a temperature-sensitive EcoRI methylase was cotransformed with different plasmids containing inserts, $(G)_9AATTC(G)_9$ and $(GAA)_9TTC(GAA)_8$, that are able to form intramolecular triplexes in vitro. Inhibition of methylation in vivo was found for $(G)_9AATTC(G)_9$ and $(GAA)_9TTC(GAA)_8$, suggesting that the pur pyr sequences adopt unusual strucures in E. coli. In addition, experiments using two dimensional gel electrophoresis confirmed that intramolecular triplexes are formed for the pur pyr sequences under negative supercoiling. These results demonstrate the existence of intramolecular triplexes in E. coli.

• Microbiology and Biotechnology Letters
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• v.18 no.1
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• pp.89-93
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• 1990

To improve the stability of recombinant pBR322-trip$^+$ plasmid (pLTW24) in E. coli culture, a positive selection system was devised. A DNA fragment containing pheW$^+$ gene (a structural gene for tRNA$^{phe}$ was isolated and inserted into the pBR322-trip$^+$ plasmid(pLTP24). A temperature sensitive host strain. LC901-pheS$^{-ts}$, was constructed for this plasmid by transducing pheS$^{-ts}$ allele (phenylalanyl-tRNA synthetase) to E. coli LC901 using P1kc bacteriophage. The LC901-pheS$^{-ts}$ cells were unable to grow at a restrictive temperature when they had lost the pBR322 :: pheW$^+$ (pLTP24) plasmid. The effects of pheW$^+$ gene on the plasmid stability and the expression level of trip$^+$ gene in LC901-pheS$^{-ts}$ strain were investigated. The proportion of Trip$^+$ colonies among LC901-pheS$^{-ts}$ strain carrying plasmid pLTP24 was 99%, whereas that of LC901 strain carrying plasmid pLTW24 was 7% at the end of 20 generations. After 100 generations of growth, the strain LC901-pheS$^{-ts}$ carrying plasmid pLTP24 showed little loss of plasmids. While the majority of plasmid pLTW24 in LC901 strain were lost in the same period. The activities of tryptophan synthetase (T. Sase) and anthranilate synthetase (A. Sase) in LC901 strain carrying pLTW24 were about 1.2 times and 1.8 times respectively of those in LC901-pheS$^{-ts}$ strain carrying pLTP24.

• Korean Journal of Microbiology
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• v.48 no.1
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• pp.22-28
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• 2012

Vibrio parahaemolyticus is one of the major agents responsible for food poisoning during summer in Korea, which is transmitted via seawater or seafoods. Recently, distribution of the bacteria in the marine environment has been increased due to global warming. Great concern also has been raised regarding public hygiene as well as marine culture by the emergence of pathogens with antibiotic resistance. Therefore, distribution of V. parahaemolyticus and antibiotic resistance of the isolates were monitored in 7 coastal areas of Kyonggi Province and Incheon by sampling seawater, fishes and clams monthly. V. parahaemolyticus was detected from 47.7% of 966 samples (seawater 61.9%, seafoods 41.8%) analyzed using $CHROMagar^{TM}$ and TCBS agar plates as well as multiplex PCR. Among 13 antibiotics tested, resistance to vancomycin and ampicillin was observed in 97.3% and 87.3% of the isolates, respectively, and the ratios of them resistant to cephalothin (48.8%) and rifampin (46.1%) were also high. The isolates were most highly sensitive to chloramphenicol (91.7%) and trimethoprim-sulfamethoxazole (91.8%). The ratio of sensitivity for other antibiotics was also high in the descending order of gentamycin (82.3%), tobramycin (74.8%), nalidixic acid (71.6%), tetracyclin (69.4%), cefotaxime (63.0%). About 69% of the isolates showed multiple drug resistance toward 3 antibiotics including vancomycin and ampicillin. Two of them exhibited resistance for 11 antibiotics used in this study. Plasmid profile analysis of the isolates with antibiotic resistance revealed that 55.1% of them retained plasmids of 24 different types. However, no clear inter-relationship between the resistance and the plasmid profile has been observed.

• Korean Journal of Microbiology
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• v.54 no.1
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• pp.53-59
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• 2018

The objective of this study was to investigate the plasmid profiling of multi-drug resistant (MDR) Vibrio in influent (inflow) and effluent (discharged) water samples of fish farms in Jeju, South Korea. MDR isolates identified through disc diffusion susceptibility tests, were subjected to plasmid profiling. One hundred fifty Vibrio isolates were obtained from each influent and effluent water sample. All MDR isolates were subjected to plasmid profiling. Greater number of bacteria were enumerated from effluents (61%) comparing to influents (39%). High incidence of neomycin, sulfamethoxazole, amoxicillin and oxytetracycline resistance was observed among the isolates, which was higher in effluent samples. In contrast, Vibrio isolates were more susceptible to florfenicol, chloramphenicol, ciprofloxacin, and nalidixic acid. Among 99 (influent 39 and effluent 60) MDR isolates, a total of 58 (influent 38 and effluent 20) were found to bear plasmids ranging from 1.7 kb to >10 kb and showed 19 different antibiograms according to the size of plasmids. MDR isolates showed six and four distinct plasmid profiles in influent and effluent, respectively. Effluent samples contained more plasmid-carrying MDR Vibrio isolates with more diverse plasmid profiles and antibiograms, suggesting that fish farm tanks may serve as a reservoir of antibiotic resistance genes. The presence of plasmid-carrying MDR Vibrio isolates in fish farm effluent water may contribute to the dissemination of antibiotic resistance genes to the environments, which ultimately poses threat to human health.

• Korean Journal of Microbiology
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• v.54 no.3
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• pp.266-271
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• 2018

Effect of plasmid curing of Acinetobacter sp. B-W on the production of siderophore from glutamic acid as both carbon and nitrogen sole sources was investigated. Plasmid cured mutant of strain B-W lost the ability to produce siderophore from glutamic acid at $28^{\circ}C$. Transformant E. coli $DH5{\alpha}$ harboring 20 kb plasmid, that was isolated from wild type of strain B-W produced siderophore from glutamic acid as both carbon and nitrogen sole sources at $28^{\circ}C$, but, not at $36^{\circ}C$. Production of siderophore from glutamic acid by transformant E. coli $DH5{\alpha}$ was completely inhibited by $10{\mu}M\;FeCl_3$. In previous report, catechol nature of siderophore produced from glutamic acid by strain B-W was detected by Arnow test. The siderophore produced from glutamic acid by transformant E. coli $DH5{\alpha}$ was also catechol type. Rf value of siderophore produced from transformant E. coli $DH5{\alpha}$ grown in medium glutamic acid as both carbon and nitrogen sole sources at $28^{\circ}C$ was 0.32 in butanol-acetic acid-water (12:3:5) as developing solvent. Rf value of the siderophore was the same with that of wild type of strain B-W. Thus a single plasmid of 20 kb seemed to be involved in the production of siderophore from glutamic acid.

• Proceedings of the Korean Society of Life Science Conference
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• 2005.09a
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• pp.120-120
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• 2005

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1976.10a
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• pp.185-186
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• 1976

현재 분자생물학계에서 가장 관심을 모으고 있는 문제는 한 생물체에서 DNA의 토막을 짤라 그것을 파아지 플라스미드 DNADML 토막에 붙여서 대장균이나 혹은 다른 생물체에 삽입하므로써 특정한 유전정보를 한 종으로부터 다른 종으로 이전시키는 실험들이다.(중략)

• 대한치주과학회:학술대회논문집
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• 2003.11a
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• pp.37-38
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• 2003