• 대한수의학회지
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• 제33권1호
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• pp.81-85
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• 1993

Gel electrophoresis has proven to be one of the most useful of DNA separation and purification. The new technique of pulsed field gel electrophoresis (PFGE) is high resolution separation of large size DNA moleculs. Conventional continuous gel electrophresis can not be separation of large DNA fragments(20~50 k base). Field inversion gel electrophoresis(FIGE) is very useful for large DNA molecules. We have found that a pulse ratio ; 2 : 1, time ; 24hrs., volts ; $10^{volts}/_{cm}$, start ; 0.45sec, end ; 1sec, is most effectively resolves DNA fragment in the 6~50k base.

• Journal of Applied Biological Chemistry
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• 제48권2호
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• pp.58-64
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• 2005

Listeria monocytogenes is a high-risk foodborne pathogen responsible for foodborne listeriosis outbreaks, and is particularly dangerous to immuno-compromised people with mortality rate of about 30%. This review summarizes subtyping of L. monocytogenes using Pulsed Field Gel Electrophoresis, widely used to trace origin of foodborne outbreaks and to determine relationship between isolates.

• Journal of Microbiology
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• 제35권3호
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• pp.172-176
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• 1997

The Mycobacterium tuberculosis clinical isolates in Korea, showing different drug resistances, were analyzed by comparing large restriction fragment (LRF) patterns produced y digestion of genomic DNA with infrequent-cutting endonucleases of SpeI, AsnI and pulsed-field gel electrophoresis (PFGE). SpeI and AsnI allowed with AsnI and SpeI, strains yielded an absolutely identical pattern for Korean type's mycobacteria even though they showed different drug resisstance. However, when three M. tuberculosis strains, showing drug resistance, were digested with XbaI, patterns were different from those of the other M. tuberculosis strians which are susceptible to drugs. This stuyd reveals that the comparison of chromosomal restriction patterns is very useful as an additional aid for the differentiation and identification of M. tuberculosis strains showing drug resistances.

• Journal of Microbiology and Biotechnology
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• 제16권11호
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• pp.1814-1818
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• 2006

Vibrio cholerae O1 El Tor isolates (n=242), collected in Korea between 1991 and 2002, were classified by NotI-digested pulsed-field gel electrophoresis (PFGE). The major types were Al before 1998 and B after 1999, among both domestic and imported cases. The prevalent PFGE types among domestic cases were consistent with the prevalent types among imported cases in the same year. These results suggest a close relationship between the domestic and imported cases of cholera in Korea.

• Journal of Microbiology and Biotechnology
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• 제5권1호
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• pp.1-5
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• 1995

A genomic DNA of alkaliphilic bacterium, Micrococcus sp. Y-l, was analysed using the physical mapping method of pulsed-field gel electrophoresis (PFGE). Five restriction enzymes of Sspl, Hpal, Xbal, Ndel or EcoRI, which recognize the Adenine-Thymine-rich sequences of genomic DNA, were used for the generation of few (7 to 20) distinctly separate fragments, with average sizes in the range of 200～500 kb. However, the sites for Notl and SfiI, 8 base-recognizing enzymes, were highly frequent. The genome size of this strain was determined to be 4 mega base pairs (Mb) from restriction fragments separated by PFGE. This is the first case of restriction mapping in alkaliphilic bacterium.

• 대한의생명과학회지
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• 제11권1호
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• pp.9-13
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• 2005

A total of 22 Salmonella enterica serotype Enteritidis (S. Enteritidis) isolates from human and chicken sources were analyzed by pulsed field gel electrophoresis (PFGE) using XbaI restriction enzyme to assess the genetic relationships between strains from different sources. PFGE permitted the resolution of XbaI restriction fragments of the 22 S. Enteritidis into 6 distinct PFGE types (PFT), designated PFT1 to PFT6, and 2 subtypes within PFT2, and allowed to detect between 9 and 10 bands with fragments sizes in the range of $25\~635\;kb$. Four of twelve isolates from human showed an identical PFGE patterns with 2 isolates from chickens. Also, another one isolate from human showed an identical PFGE patterns with other 5 isolates from chickens. Only one isolate from chicken, however, showed a different pattern compared to other PFTs. These results suggested that sporadic human food-poisoning cases infections caused by S. Enteritidis in this study were due to the consumption of contaminated chicken meats and that a clonally highly similar strains exist and spread between human and chicken sources.

• 대한수의학회지
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• 제39권4호
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• pp.811-818
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• 1999

A total of 16 Staphylococcus epidermidis isolates collected from 14 dogs admitted to the Veterinary Medicial Teaching Hospital in Seoul National University over eleven months were examined for in vitro antibiotic susceptibility pattern with minimum inhibitory concentration (MIC) and slime production, a virulence-associated phenotype, and were genetically characterized by pulsed-field gel electrophoresis (PFGE). The frequency of resistance to antimicrobial agents tested was not high, with a susceptibility ranging from 56.3% to 100%. Three strains exhibited multiple drug resistance against amikacin (MIC, $32-64{\mu}g/ml$), ampicillin ($32{\mu}g/ml$), fosfomycin ($32-128{\mu}g/ml$) and gentamicin ($16{\mu}g/ml$). Vancomycin, ciprofloxacin and rifampin were effective antibiotics against the isolates. All isolates were slime producers ; strains isolated from dogs which died of bacteremia were more likely to produce slime than those isolated from dogs which survived. Chromosomal DNA fingerprinting of the isolates yielded 16 different genomic types with few common bands, indicating a variety of clones of S epidermidis were prevalent in the hospital. This study revealed that PFGE is an useful method for the genotype characterization of S epidermidis strains and this organism could probably be pathogenic in some dogs with severe disorders. Further works on a larger number of epidemiologically defined strains are required to assess these results.

• 대한수의학회지
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• 제39권2호
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• pp.338-342
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• 1999

Twenty three strains of Escherichia (E) coli O157 : H7 isolated from Korea, Japan, USA were analyzed by pulsed-field gel electrophoresis (PFGE) of XbaI-digested chromosomal DNA and multiplex polymerase chain reaction. Various PFGE patterns of E. coli O157 : H7 were found on the same farm. Most of the E, coli O157 : H7 strains had shiga-like toxin (slt) II gene only (43.5%) or both slt I and slt II genes(30.4%). eaeA gene was highly conserved in the E. coli O157 : H7. There was no correlation between PFGE and slt gene patterns. The results indicate that various genotypes of E. coli O157 : H7 have spread throughout the country and genomic DNA patterns generated by PFGE are highly specific for different strains and have significant value in epidemiologic investigations of infectious disease outbreaks.

• 식품기술
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• 제22권3호
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• pp.502-512
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• 2009

• 한국균학회지
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• 제27권2호통권89호
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• pp.112-118
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• 1999

한국, 일본 그리고 미국 등지에서 수집된 Fusarium oxysporum f. sp. lycopersici의 electrophoretic karyotype(EK)을 분석하고자 CHEF-DRII pulsed field gel electrophoresis system(Bio-Rad Laboratories, Melville, NY)으로 각 공시균의 chromosome sized DNA를 분리하였다. EK 분석에 적합한 CHEF gel electrophoresis 조건을 얻기 위해 전기영동 시간 및 전압 그리고 switching interval 등의 조건을 다양하게 바꾸어 가며 실험하였다. 그 결과 국내 균주에서 $0.76{\sim}6.41\;Mb$에 달하는 $9{\sim}11$개의 chromosome sized DNA가 분리되었으며 그 total genome size는 $35.29{\sim}38.92\;Mb$ 이었다. 또한 일본과 미국 균주로 부터 $1.24{\sim}6.85\;Mb$범위의 $9{\sim}11$개의 chromosome sized DNA가 분리되었고 그 total genome size는 $35.32{\sim}43.87\;Mb$ 이었다. 이와 같이 얻어진 각 공시균주의 EK는 chromosome sized DNA의 length range 및 total genome size에서 국내 균주와 외국 균주간의 차이를 잘 반영하였다. 또한 국내 균주의 chromosomal polymorphism은 그 변이가 적어 서로 동일하거나 유사하였으며 외국 균주와 뚜렷이 다른 chromosomal DNA pattern을 나타냈다.