• 대한수의학회지
• /
• 제48권3호
• /
• pp.275-285
• /
• 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.

• 한국수정란이식학회지
• /
• 제17권2호
• /
• pp.109-115
• /
• 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.

• 한국수정란이식학회지
• /
• 제17권2호
• /
• pp.101-108
• /
• 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.

• 한국미생물·생명공학회지
• /
• 제17권5호
• /
• pp.447-453
• /
• 1989

항생물질 내성유전자를 유전자 조작기법으로 재조합시켜 만든 세균에서 그 유전자들이 전이되는 특성을 하폐수의 자연환경에서 연구하기 위하여, 자연계로부터 분리한 Gram 음성세균 중에서 kanamycin (Km), chloramphenicol (Cm), streptomycin (Sm), sulfadiazine(Su)에 내성을 띄는 균주로 DK1을 선발하였다. DK1 균주는 4개의 plasmid를 가지고 있으나 그 중 크기가 약 68kb인 pDK101 plasmid에 Km$^{${\gamma}$}$ Cm$^{${\gamma}$}$ 유전자를 가지고 있었다. 이 pDK101 plasmid에는 EcoRI site가 16개, PstI site가 32개, SalI site가 6개씩 있었다. pDK101 plasmid 와 vector로 사용한 pKT230을 E. coli으로 처리하여 얻은 절편들로부터 Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ 유전자를 가지는 pDT309와 pDTS29 등의 chimeric piasmid를 제조하였다. 이들을 다시 E. coli C600과 E. coli HB101 에 형질전환 시킴으로써, DKC601, DKC602, DKH 102 그리고 DKH103 등의 재조합 균주를 얻었다. 이들 재조합 균주에서는 모두 Km$^{${\gamma}$}$Cm$^{${\gamma}$}$유전자가 잘 발현되었으며, 그 중 DKC601과 DKH103에서는 각각 pDT529와 pDT309의 chimeric plasmid가 포함되어 있는 것이 전기영동 방법으로 명확히 확인되었다.

• Journal of Microbiology and Biotechnology
• /
• 제28권2호
• /
• pp.330-337
• /
• 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.

• 약학회지
• /
• 제35권2호
• /
• pp.61-72
• /
• 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
• /
• 제45권3호
• /
• pp.193-198
• /
• 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.

• 미생물학회지
• /
• 제23권2호
• /
• pp.138-146
• /
• 1985

포도당 이성화효소를 coding는 Bacillus licheniformis ATCC31667의 유전자를 Escherichia coli LE 392-6에 클로닝하였다. Bacillus lieheniformis 염색체 DNA를 분리하고 제한효소인 Pst I.HindIII, Sal 1, EcoR 1, BamH1으로 절단한 후 운반제 plasmid인 pBR332에 연결하고 포도당 이성화효소 negative인 E. coli LE 3926-6에 형질전환하였다. 이중 E채꺄 제한효소를 사용한 것만이 glucose isomerase positive로 전환되어 xylose를 유일 탄소원으로 하여 성장하였다. 이 제조합 plasmid를 제한효소로 처리하여 본 결과 4.1Kb의 Bacillus licheniformisdb전자가 옮겨 졌음을 확인했고 여기에 제한효소 HindII와 Puv II의 절단위치가 확인되어 제한요소 지도를 작정하였다. 이 재조합 plasmid pBGI6는 연속계대 10일 후에도 매우안정하게 유지되었다. 한편 포도당 이정화 효소의 안정을 측정하여 본 바 야생숙주에 비해 약 20배의 증가를 나타냈다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제7권3호
• /
• pp.427-434
• /
• 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.

• 미생물학회지
• /
• 제28권3호
• /
• pp.188-191
• /
• 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.