• 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.

• Journal of Embryo Transfer
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• v.17 no.2
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• pp.109-115
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• 2002

The present study was conducted for the production of transgenic cloned cows those secrete human lactoferricin into milk by somatic cell nuclear transfer (NT). To estimate detrimental effects of gene transfection on transgenic cloned embryo production, development rates of NT embryos were compared between transfected and non-transfected cumulus and ear fibroblast cells. An expression plasmid for human lactofericin (pbeta-LFC) was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and human lactoferricin target gene into a pcDNA3 plasmid. Two bovine somatic cell lines (cumulus cell and ear fibroblast) were established and transfected with the expression plasmid using a liposomal transfection reagent, Fugene6 as a carrier. Cumulus cell and ear fibroblast were transfected at the passage of 2 to 4, trypsinized and GFP-expressing cells were randomly selected and used for somatic cell NT. Developmental competences (rates of fusion, cleavage, and blastocyst formation) in bovine transgenic somatic cell NT embryos reconstructed with non-transfectecd cells were significantly higher than those from transfected cells in cumulus cell and ear fibroblast (P＜0.05). This study indicated that transfection of done. cell has detrimental effect on embryo development in bovine transgenic NT.

• Journal of Embryo Transfer
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• v.17 no.2
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• pp.101-108
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• 2002

Human Prourokinase (proUK) offers potential as a novel agent with improved fibrin specificity and, as such, may offer advantages as an attractive alternative to urokinase that is associated with clinical benefits in patients with acute peripheral arterial occlusion. For production of transgenic cow as human proUK bioreacotor, we conducted this study to establish efficient production system for bovine transgenic embryos by somatic cell nuclear transfer (NT) using human prourokinase gene transfected donor cell. An expression plasmid for human prourokinase was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and human prourokinase target gene into a pcDNA3 plasmid. Cumulus cells were used as donor cell and transfected with the expression plasmid using the Fugene 6 as a carrier. To increase the efficiency for the production of transgenic NT, development rates were compared between non-transfected and transfected cell in experiment 1, and in experiment 2, development rates were compared according to level of GFP expression in donor cells. In experiment 1, development rates of non-transgenic NT embryos were significantly higher than transgenic NT embryos (43.3 vs. 28.4％). In experiment 2, there were no significant differences in fusion rates (85.4 vs. 78.9％) and cleavage rates (78.7 vs. 84.4％) between low and high expressed cells. However, development rates to blastocyst were higher in low expressed cells (17.0 vs. 33.3％), and GFP expression rates in blastocyst were higher in high expressed cells (75.0 vs. 43.3％), significantly.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.447-453
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• 1989

In order to study the transfer of antibiotics resistance genes of the genetically cloned bacteria in water environments, DK1 strain, which is resistant to kanamycin (Km), chloramphenicol (Cm), streptomycin (Sm), and sulfadiazine (Su), was selected from the Gram-negative bacterial isolates from wastewater. One of 4 plasmids harboured in the DK1 strain was found to possess Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ gene and be about 68 kb in size, and it was designated as pDK101. The plasmid of pDK101 was also found to have 16, 32, and 6 restriction sites for EcoRI. .PstI, and SalI, respectively. From the digestion fragments of pDK101 plasmid and pKT230 used as a vector by EcoRI restriction endonuclease, pDT309 and pDT529 were constructed as chimeric plasmids which possess Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ gene and are 30.9 and 52.9 kb in size, respectively. When the chimeric plasmids were trasformed into E. coli C600 or E. coli HB101, transformants of DKC601, DKC602, DKH102, and DKH103 were obtained as cloned bacterial cells. The Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ genes were well expressed in those cloned cells and the chimeric plasmids were clearly detected in the cloned cells of DKC601 and DKH103.

• Journal of Microbiology and Biotechnology
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• v.28 no.2
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• pp.330-337
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• 2018

Polycyclic aromatic hydrocarbons (PAHs), including naphthalene, are widely distributed in nature. Naphthalene has been regarded as a model PAH compound for investigating the mechanisms of bacterial PAH biodegradation. Pseudomonas sp. AS1 isolated from an arseniccontaminated site is capable of growing on various aromatic compounds such as naphthalene, salicylate, and catechol, but not on gentisate. The genome of strain AS1 consists of a 6,126,864 bp circular chromosome and the 81,841 bp circular plasmid pAS1. Pseudomonas sp. AS1 has multiple dioxygenases and related enzymes involved in the degradation of aromatic compounds, which might contribute to the metabolic versatility of this isolate. The pAS1 plasmid exhibits extremely high similarity in size and sequences to the well-known naphthalene-degrading plasmid pDTG1 in Pseudomonas putida strain NCIB 9816-4. Two gene clusters involved in the naphthalene degradation pathway were identified on pAS1. The expression of several nah genes on the plasmid was upregulated by more than 2-fold when naphthalene was used as a sole carbon source. Strains have been isolated at different times and places with different characteristics, but similar genes involved in the degradation of aromatic compounds have been identified on their plasmids, which suggests that the transmissibility of the plasmids might play an important role in the adaptation of the microorganisms to mineralize the compounds.

• YAKHAK HOEJI
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• v.35 no.2
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• pp.61-72
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• 1991

These studies were made to assess the present stage of resistance to antibiotics, incidence and transferability of R-factors against E. coli. From March to July 1987, 59 strains of E. coli were isolated from specimens of patients collected at university hospitals in Seoul, 64 strains from stools of domestic animals and 66 strains from drainages in Seoul. These specimens were tested for resistance to 12 kinds of antimicrobial agents by means of agar dilution method. Using Muller-Hinton agar for the assay of drug resistance and tryptic soy broth as propagating medium for conjugation. The strains of E. coli were found to be resistant to one or more antibiotics and were considered to be potential donors of R-plasmid. The resistant strains of E. coli isolated from patients, domestic animals and drainages were found to be 55(93%), 33(52%) and 31(47%), respectively. Resistance to Tc, Ap and Cb was the highest in those isolated from patients and drainages, and resistance to Tc, Cm and Sm was the highest in those isolated from domestic animals. In the transfer test of drug resistance by conjugation method, 17 strains (47%) isolated from patients, 15(54%) isolated from domestic animals and 15(56%) isolated from drainages showed positive results, transperable resistant plasmid molecules with variable range in each strain.

• Journal of Microbiology
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• v.45 no.3
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• pp.193-198
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• 2007

A marine bacterium was isolated from Mai Po Nature Reserve of Hong Kong and identified as Vibrio cholerae MP-1. It contains a small plasmid designated as pVC of 3.8 kb. Four open reading frames (ORFs) are identified on the plasmid, but none of them shows homology to any known protein. Database search indicated that a 440 bp fragment is 96% identical to a fragment found in a small plasmid of another V. cholerae. Further experiments demonstrated that a 2.3 kb EcoRI fragment containing the complete ORF1, partial ORF4 and their intergenic region could self-replicate. Additional analyses revealed that sequence upstream of ORF1 showed the features characteristic of theta type replicons. Protein encoded by ORF1 has two characteristic motifs existed in most replication initiator proteins (Rep): the leucine zipper (LZ) motif located at the N-terminal region and the alpha helix-turn-alpha helix motif (HTH) located at the C-terminal end. The results suggest that pVC replicates via the theta type mechanism and is likely a novel type of theta replicon.

• Korean Journal of Microbiology
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• v.23 no.2
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• pp.138-146
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• 1985

A Bacillus licheniformis ATCC31667 gene coding for a glucose isomerase has been cloned and expressed in glucose isomerase negative mutant of Escherichia coli. A recombinant plasmid, constructed by ligation of a EcoRI fragment of B.licheniformis chromosomal DNA to vector plasmid pBR322, was expressed glucose isomerase positive in E.coli LE392-6 with growth on minimal medium containing xylose as a sole carbon source. This recombinant plasmid, designated pBGI6, had the insery of 4.1Kb of Bacillus gene in EcoRI site, and restriction map of the plasmid was established. The plasmid pBG16 was very stable after 10days of serial transfer to a fresh medium. The activity of glucose isomerase from the transformed cell containing pBGI6 was increased about 20 fold than its wild type of host.

• Asian-Australasian Journal of Animal Sciences
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• v.7 no.3
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• pp.427-434
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• 1994

The primordial germ cells (PGCs) were transfected in vitro and expressed the exogenous RSVLTR/${\beta}G2$ plasmid, suggesting thaI PGC is a possible vector for direct gene transfer into the germ line. Transfection efficiency of cell suspensions containing PGCs was 1.5% by liposome mediated DNA transfection. By microinjection of the transfected PGCs into the host germinal crescent, PGCs migrated via blood vessel to the future gonad and these transfected PGCs resulted in the RSVLTR/${\beta}G2$ expression in the gonad. The results from the seeding of PGCs on the chorioallantoic membrane were insufficient to test the hypothesis that PGCs can penetrate or invade the chorioallantoic membrane for transport via the circulatory system.

• Korean Journal of Microbiology
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• v.28 no.3
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• pp.188-191
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• 1990

The streptococcal plasmid pAM$\beta_{1}$ (erythromycin resistance)was transferred via conjugation from Streptococcus faecalis DS 5 to Lactobacillus casei YIT 9018 by a filter mating method. The transfer frequency depended greatly on the type, pore size and side(front or back) of membrane filter. Water passing through a membrane under reduced pressure induced very tight contact between the cells, increased the transconjugation frequency about 1.3 to 37-fold when Millipore membrane filter (0.45.$\mu$m, front side up) was used.