• Korean Journal of Veterinary Research
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• v.30 no.3
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• pp.287-295
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• 1990

The purpose of this study was the examination for presence of pilus antigen, O serogroups, colicin production, antibiotic susceptibility and plasmid profiles among E coli isolated from diarrheal piglets and fattening pigs in Taegu province. Of 145 E coli isolated, 98 strains (67.4%) possesed pilus antigens which belonged to either K88 (47.6%), K99 (11.7%) or 987P (8.3%) types. Fifty-nine strains (40.7%) were classified into tenO serogroups and their types were O8 (22.0%), O20(16.9%), O141(15.3%), O9(10.2%), O45(10.2%), O139(8.5%), O064(6.8%), O149(5.0%), O157(3.4%), and O115(1.7%). Thirty-three strains (22.8%) were colicinogenic and 6 strains (4.1%) were hemolytic. One hundred and thirty-nine strains (95.9%) of 145 E coli isolates were resistant to ampicillin, chloramphenicol, gentamicin, kanamycin, streptomycin, tetracycline, rifampicin and nalidixic acid, alone or in combination thereof. Ninety strains (64.7%) of 139 drug resistant strains carried R factor (R) which were transferable to the recipient by conjugation. In gel electrophoresis for the isolation of plasmid DNA, the number of plasmid DNA band varied from 2 to 11 in 16 E coli with pilus antigen. It's molecular weight ranged from 1.0 to 60.0 megadalton.

• Journal of Microbiology and Biotechnology
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• v.3 no.4
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• pp.256-260
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• 1993

DNA subfragments, sopA, sopB and sopC which help to maintain the stability of an ori C plasmid, were derived from a mini-F plasmid DNA (EcoRI restriction fragment f5) after digestion with restriction endonuclease, and cloned in the vector plasmid pBR322. The recombinant plasmids obtained were introduced into E. coli KY7231 and E. coli CSR603 strains, and proteins specified by the mini-F fragments were analysed by SDS-PAGE. Two proteins encoded by the F fragments were detected, and their molecular weights were 41,000 and 37,000 daltons. Fluorography after one and two dimensional gel electrophoresis of the lysates showed that these two proteins had been overproduced in the cells which were allowed to incorporate radioactive amino acid after plasmid amplification by chloramphenicol treatment. The isoelectric points of sopA and sopB proteins were 6.6 and 7.0, respectively.

• Korean Journal of Veterinary Research
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• v.34 no.4
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• pp.759-768
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• 1994

E coli strains isolated from pigs were investigated with respect to antimicrobial resistances and prevalence of R-plasmids. Also determined were properties of R-plasmids by plasmid conjugation, curing and southern hybridization using gene probes. All of 400 E coli strains were resistant to CL and SU, and 0.3% to 96.8% of the strains were resistant to most antimicrobials such as TC, PG, AM, SM, CP, GM, EM, NM, etc, while all strains were sensitive to AK. All strains were also multiply resistant to three to twelve antimicrobials. The resistances to PG, SM, TC, AM, CP, SU and ST were transferable and supposed to be mediated by R-plasmids which were opportunistic for transposition into chromosome. Plasmids bigger in size than chromosomal DNA were considered as R-plasmids and most plasmids in small size (<4Kb) proved as cryptic plasmids or nonconjugative R-plasmids. In a strain(No 99), AM resistant property was determined from both chromosomal DNA and R-plasmid DNA which is bigger in size than chromosome.

• Korean Journal of Microbiology
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• v.43 no.2
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• pp.116-123
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• 2007

The purpose of this study was typing the plasmid mediated extended spectrum ${\beta}-lactamases$ produced by enteric bacteria isolated from rivers in Pusan. Six strains of Eschericha coli and fifteen strains of Klebsiella pneumoniae transferred their plasmid mediated extended spectrum ${\beta}-lactamase$ genes to the recipient strain Eschericha coli J53 $Azid^{R}$. The plasmid mediated extended spectrum ${\beta}-lactamase$ genes were sequenced directly after PCR and the types were determined by the BCM Search Launcher and GenBank nucleotid database. Determined types of the plasmid mediated extended spectrum ${\beta}-lactamases$ were TEM-52 and SHV-12. TEM-52 was isolated from both Escherichia coli and Klebsiella pneumoniae. However SHV-12 was isolated from Klebsiella pneumoniae only. The results indicated that the plasmid mediated extended spectrum ${\beta}-lactamase$ producing bacteria spreded over the area of clinical to the nature in Korea.

• Journal of Microbiology and Biotechnology
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• v.5 no.6
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• pp.370-372
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• 1995

A Phaffia rhodozyma chromosomal fragment (approximately 3.8 kb) capable of functioning as an origin for the replication of a kanamycin resistance ($Km^r$) plasmid in S. cerevisiae was isolated by the use of origin search plasmid, pHN134. In S. cerevisiae, transformation frequencies using the plasmid pHN134 containing an autonomously replicating sequence of P. rhodozyma was 450-580 CFU/$\mu g$ DNA. The stability of the recombinant plasmid were 16-19$\%$.

• Korean Journal of Veterinary Research
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• v.48 no.3
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• pp.275-285
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• 2008

Antimicrobial drugs are widely used in poultry industry as growth promoters or to control infectious diseases. However, this practice is reported to have caused high resistance to antimicrobial drugs in normal chicken flora and pathogens. Antimicrobial resistance to Escherichia coli (E. coli) from chicken has been mainly reported in normal flora, but rare in pathogenic organism in Korea, recently. Therefore, this study was conducted to investigate prevalence of antimicrobials resistance, transfer of R plasmid, and association between antimicrobial drug resistance and O serotype of 203 pathogenic E. coli from poultry in Korea during the period from April 2003 to December 2005. These isolates showed a high resistance to tetracycline (Tc, 93.6%), nalidixic acid (Na, 92.6%), streptomycin (Sm, 81.8%), ampicillin (Ap, 77.3%), ciprofloxacin (Ci, 70.9%), sulfisoxazole (Su, 66.5%), and trimethoprim (Tp, 58.1%). Two hundred-one (99.0%) of the isolates were resistant to one or more drugs. They showed 57 different resistant patterns, and the most prevalent resistant pattern among them was Tc, Sin, Su, Ap, Tp, Ci, Na. Sixty-eight (33.8%) of the isolates transferred all or a part of their antimicrobial resistant pattern to the recipient strain by R plasmid. The most common antimicrobial resistant pattern was Tc, Sm, Su, Ap, Tp, Ci, Na in serotype O78, O88 and O15, respectively. These results exhibit high individual and multiple resistance to antimicrobials of pathogenic E. coli from poultry in Korea. They also suggest the needs for surveillance to monitor antimicrobial resistance in pathogenic bacteria that can be potentially transmitted to humans from food animals and to regulate the abuse of antimicrobials on food-producing animals in Korea.

• Korean Journal of Microbiology
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• v.30 no.3
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• pp.218-224
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• 1992

Recombinant plasmid shuttle vectors were constructed for genetic studies on the oxidation of carbon monoxide by carboxydobacteria. Two vectors. pYK322 (7.2 kb, Ap'. Tc') and pYK 324 (7.2 kb, Ap', Tc'), were constructed using pBR322 and pYK100. a small plasmid in Pseudomonas carbo,xydovorans. Four plasmids. pYK2IO (5.2 kb. Cm'), pYK220 (5.2 kb, Cmr), pYK230 (5.2 kb, Cm'), and pYK232 (5.2 kb. Cm'), were constructed using pACYC184 and pYK100. Transformation of several carboxydobacteria with pYK322 and pYK220 was round to be efficient when the cells were transformed by the methoti of Bagdasarian and Timmis (Curr. Top. Microbiol. Immunol. 96:47-67. 1982) with several modifications; cells growing on 0.2% succinate were harvested at the mid-exponential phase. 10 mM RbCl in transformation solution was substituted with 100 mM KCI. cclls in transformation solution were incubated for 12 h at 4'C before addition of DNA and heat shock was carried out for 3 min at 45$^{\circ}$C. Plasmid vectors used for transformation, however. were not detected from antibiotics-resistant transformants, suggesting that the vectors may be integrated into the chromosomal DNA.

• The Journal of the Korean Society for Microbiology
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• v.22 no.1
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• pp.35-53
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• 1987

Multiply resistant Shigella strains isolated in Taegu area were subjected for the characterization of R plasmids. All strains isolated in 1984 and 1985 were susceptible to gentamicin, amikacin, and cephalothin, and most strains were susceptible to kanamycin (Km) and rifampin by agar dilution antimicrobial susceptibility test. The resistance frequency of S. flexneri against ampicillin (Ap) was higher than that of S. sonnei. The strains resistant to sulfisomidine (Su) and trimethoprim (Tp) were found at higher frequency in S. sonnei than in S. flexneri. The most prevalent resistance pattern of S. flexneri was chloramphenicol (Cm) tetracycline (Tc) streptomycin (Sm) Ap, followed by the pattern of CmTcSmSuApTp, CmTcSmSuApTp nalidixic acid, and CmTcSmSuAp in the decreasing order. The antibiogram of CmTcSmSuTp was found to be the most frequent pattern in S. sonnei. The ratio of conjugal transfer of S. flexneri was 47% and 75% of S. sonnei. The average number of plasmid harboring in Shigella was 4 and the size of plasmid ranged 1.3 to 134 megadalton (Mdal). Most S. flexneri carried plasmids of 2 to 3 Mdal and S. sonnei carried those of 3 to 4 Mdal size. The sizes of conjugative plasmids ranged 40-90 Mdal. The incompatibility group (Inc) F II plasmids (54-59 Mdal) were most frequent and rare Inc B plasmids (60 Mdal) of isolates in 1979 and 1980 and Inc FI (87 Mdal) of 1983 isolates were able to be classified by the colony test with standard reference plasmids. The R plasmids of known Inc group were tested for the restriction endonuclease analysis. The pattern of plasmids digested by EcoRl were apparently different by the Inc group but there was no significant difference between species or by the resistance patterns. Nonconjugative plasmids and their phenotypes were identified by transformation test. The transformants were resistant to less than two drugs. Colicin producing transformants carried the Col plasmid of 3.7 or 3.9 Mdal size. $Ap^r$ plasmids derived from S. sonnei were found to be mobilized by transfer factor RT641 to E. coli #CS100. $Ap^r$ plasm ids of same size shared by S. flexneri, S. sonnei, and E. coli were digested with Pstl. All of them showed two restriction fragments of 2.8 kilobase(kb) and 0.7kb. Other plasmids ($Sm^r\;Su^r$) derived from S. flexneri, S. boydii, and S. sonnei were digested with Pstl and they showed same restriction fragment patterns of 3.1kb and 2.9kb. The plasmid profiles of three strains of S. sonnei producing colicin and showing same resistance pattern of CmTcSmSuApTpKm appeared to be similar. Restriction patterns by EcoRl and the behavior of plasmids in conjugation or transformation process were also similar between those plasmids. The restriction patterns were significantly different between the plasmids of Inc FI group and those of unclassified Inc group.

• Korean Journal of Microbiology
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• v.25 no.4
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• pp.262-273
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• 1987

The replication region of the chloramphenical resistance plasmid pSBK203 of Staphylococcus aureus was cloned using pBR322 and pBD9 as vectors. Cloned replication tegion and chloramphenicol resistance gene were recombined to pBR322. The reconstructed vector behaved as a shuttle vector for E. coli and B. subtilis.

• The Journal of the Korean Society for Microbiology
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• v.13 no.1
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• pp.7-16
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• 1978

One hundred and three clinical isolates of enterococci were examined for susceptibility to 8 antibiotics, and transferability and transfer frequency of R plasmid. Ampicillin was the most active, followed in decreasing order by rifampin, amikacin and chloramphenicol, and tetracycline. High-level resistance(${\geq}2,000{\mu}g/ml$) to kanamycin, streptomycin, and gentamicin, known as the most active of the aminoglycosides to enterococcus, was present in 26.2%, 21.4%, and 18.3% of the isolates, respectively. In the drug susceptibility of the species, S. zymogenes was the most resistant and S. durans was the most sensitive to tested antibiotics. We could observed the transferability of enterococcal R plasmid in mixed culture: among the 28 strains which showed multiple drug resistance, 17 strains transferred all or part of their resistance with $2{\times}10^{-4}-2{\times}10^{-6}%$ of transfer frequency to a plasmid-free recipient, S. faecalis strain JH 2-2.