• Title/Summary/Keyword: E. Coli

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Molecular cloning and foreign gene expression of restriction endonuclease fragments of the Hc nuclear polyhedrosis virus DNA (Hc nuclear polyhedrosis virus DNA 제한효소절편의 molecular cloning 과 외래 유전자 발현)

  • Lee, Keun-Kwang
    • Journal of fish pathology
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    • v.8 no.1
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    • pp.31-36
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    • 1995
  • Hc nuclear polyhedrosis virus DNA genome was digested with EcoRI endonuclease, these partial fragments were recombined into the pUC8 plasmid vector and transformed in E. coli JM 83 cell. The genome DNA has 24 EcoRI fragments and 12 fragments of them were subcloned. The four recombinants were named as eNP-O, eNP-Q, eNP-R and eNP-S. The expression of foregin gene of the recombinants was investigated by analysing protein patterns on the SDS-PAGE. The eNP-O, eNP-Q and eNP-R were expressed a different weight of protein as comparision with potypeptide bands of E. coli JM 83 host cell.

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Antimicrobial resistance and distribution of tetracycline resistance determinants in Escherichia coli isolated from aquatic birds (수생조류에서 분리한 대장균의 항균제 내성 및 Tetracycline 내성인자의 분포)

  • Cho, Jae-Keun;Lee, Sang-Min;Kim, Ki-Seuk
    • Korean Journal of Veterinary Research
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    • v.48 no.3
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    • pp.295-303
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    • 2008
  • One hundred and sixty nine Escherichia (E.) coli strains isolated from fecal samples of aquatic birds in Geumho river basin and Dalseong park were tested by agar dilution method to determine their susceptibility patterns to 14 antimicrobial agents. The distribution of tetracycline resistance determinants (tetA, tetB, tetC, tetD and tetE) were also examined by PCR in 76 tetracycline-resistant ($TC^r$) E. coli isolates. The high resistance was observed in tetracycline, cephalothin and ampicillin (45.0~36.7%). Resistance of E. coli isolates derived from Dalseong park to tetracycline, cephalothin, ampicillin and streptomycin (65.7~44.8%) were significantly higher than those isolated from Geumho river basin (31.4~14.7%). About seventy percent (70.4%) of the strains isolated were resistant to one or more drugs tested. Thirty (39.5%) of 76 $TC^r$ E. coli isolates which were resistant to one or more drugs transferred all or a part of their resistance patterns to the recipient strain of E.coli J53 by conjugation. All of $TC^r$ E. coli isolates contained at least one or more of 5 tet genes examined. The most common genes found in these isolates were tetA (60.6%) and followed by tetB (7.9%) and tetC (1.3%). However, tetD and tetE were not found in any of the isolates tested. Twenty one (27.6%) of $TC^r$ E. coli isolates had two determinants, tetA/tetB (20 strains), tetA/tetC (1 strain). And two strains (2.6%) contained three determinants (tetA/tetB/tetC).

Quantification of Microstructures in Mice Alveolar Bone using Micro-computed tomography (${\mu}CT$)

  • Park, Hae-Ryoung;Kim, Hyun-Jin;Park, Byung-Ju
    • International Journal of Oral Biology
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    • v.38 no.3
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    • pp.87-92
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    • 2013
  • Periodontal inflammation increases the risk of tooth loss, particularly in cases where there is an associated loss of alveolar bone and periodontal ligament (PDL). Histological and morphometric evaluation of periodontal inflammation is difficult. Especially, the lengths of the periodontal ligament and interdental alveolar bone space have not been quantified. A quantitative imaging procedure applicable to an animal model would be an important clinical study. The purpose of this study was to quantify the loss of alveolar bone and periodontal ligament by evaluation with micro-computed tomography (micro-CT). Another purpose was to investigate differences in infections with systemic E. coli LPS and TNF-${\alpha}$ on E. coli lipopolysaccharide (LPS) in loss of alveolar bone and periodontal ligament model on mice. This study showed that linear measurements of alveolar bone loss were represented with an increasing trend of the periodontal ligament length and interdental alveolar process space. The effects of systemic E. coli LPS and TNF-${\alpha}$ on an E. coli LPS-induced periodontitis mice model were investigated in this research. Loss of periodontal ligament and alveolar bone were evaluated by micro-computed tomography (micro-CT) and calculated by the two- and three dimensional microstructure morphometric parameters. Also, there was a significantly increasing trend of the interdental alveolar process space in E. coli LPS and TNF-${\alpha}$ on E. coli LPS compared to PBS. And E. coli LPS and TNF-${\alpha}$ on E. coli LPS had a slightly increasing trend of the periodontal ligament length. The increasing trend of TNF-${\alpha}$ on the LPS-induced mice model in this experiment supports the previous studies on the contribution of periodontal diseases in the pathogenesis of systemic diseases. Also, our findings offer a unique model for the study of the role of LPS-induced TNF-${\alpha}$ in systemic and chronic local inflammatory processes and inflammatory diseases. In this study, we performed rapidly quantification of the periodontal inflammatory processes and periodontal bone loss using micro-computed tomography (micro-CT) in mice.

Antibacterial Activity of Garlic Extract against Escherichia coli (마늘즙액의 대장균 생육 저해 작용)

  • Kim, Youn-Soon;Park, Kyung-Suk;Kyung, Kyu-Hang;Shim, Sun-Taek;Kim, Hyun-Ku
    • Korean Journal of Food Science and Technology
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    • v.28 no.4
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    • pp.730-735
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    • 1996
  • Antibacterial action of garlic extract against Escherichia coli was investigated. When the survival of E. cloi in tryptic soy broth (TSB) containing 50% garlic extract was compared with those of Lactobacillus plantarum, Leuconostoc mesenteroides and Staphylococcus aureus, E. coli was the most sensitive to garlic antibacterial action. When E. coli was inoculated into TSB with different concentrations of garlic extract, viable cell number decreased continuously during the test period even at 1% garlic extract. When E. coli was inoculated into pH-adjusted TSB containing 0.5% garlic extract, viable cell number of E. coli decreased continuously at initial pH of 5.2 and 6.2, while it decreased initially but increased to $8.0{\times}10^{7}\;CFU/ml at 48 hr at pH 7.2. With larger initial populations $(10^{6}\;CFU/ml), E. coli grew without apparent inhibition, while with smaller initial populations $(<10^{5}\;CFU/ml), viable cell number decreased initially but later increased. Thiol compounds like cysteine and glutathione, with free SH group (s), helped E. coli to grow or survive better in TSB with inhibitory level (5%) of garlic extract. The possibility of eliminating E. coli by using garlic extract from foods like kimchi of which garlic is one of regular ingredients is suggested.

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Relationship Between Drug Resistance and Citrate Utilizing Ability of Citrate-Utilizing Variants of Escherichia Coli from Cattle (Citrate이용대장균(利用大腸菌) 변이주(變異株)의 약제내성(藥劑耐性)과 Citrate이용성(利用性)과의 관계(關係))

  • Lee, Hun Jun;Kim, Kyo Joon;Choi, Won Pil
    • Korean Journal of Agricultural Science
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    • v.10 no.2
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    • pp.200-205
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    • 1983
  • Transfer experiment of drug resistance and citrate utilizing ability were performed to show the relationship between drug resistance and citrate utilizing ability of 76 Citrate-Utilizing variants of Escherichia coli(Cit $^+E$. coli) isolated from cattle. The results obtained were summarized as follows; 1. Of seventy-six Cit $^+E$. coli, 36(47.4%) strains transfered their drug resistance to the E. coli ML 1410 by mating. 2. Tetracycline(TC) resistance determinants and citrate utilizing ability were transmitted in the recipient strains selected for TC, but it can be seen the segregation of streptomycin (SM) resistance determinants and citrate utilizing character in 9 recipient strains selected for SM. 3. Of seventeen TC resistance Cit $^+E$. coli, 10 strains transfered citrate utilizing character, alone. 4. The transmission of the ability to utilize citrate on Simmons citrate agar at $37^{\circ}C$, was demonstrated 52(68.4%) out of the 76 Cit $^+E$. coli.

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Colony Count with Mixed Culture of Enteric Bacteria by in vitro Quantitative Method (장내세균의 시간차 혼합배양이 보여주는 균수측정의 비교)

  • 황선철;전보성
    • Korean Journal of Microbiology
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    • v.11 no.4
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    • pp.175-180
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    • 1973
  • This study was attempted to see more clear relationships among the enterobacteria, especially between the intestinal normal flora and pathogenic bacteria. It has been known that some intestinal normal flora produce the bactrial metabolites that are harmful to other enteric bacteria. One of the metabolites is known as colicin, the protein fraction, which possesses certain degree of inhibitory effect against other bacterial growth fraction, whih possesses certain degree of inhibitory effect against other bacterial growth. As a preliminary study for a colicin purification, the antagonistic effect of E, coli to groups of Salmonella and Shigella has been studied by means of in vitro quantitative culture method. 1. E.coli showed definite inhibitory effects aganist both Salmonella and Shigella groups in the mixture of two organisms. 2. The inhibitory effects of E.coli in the E.coli-Salmonella and the E.coli-Shigella mixture occurred from 4 hours incubation following the inoculation. 3. Even the complete inhibition of pathogenic enteric bacterial growth was noticed in the E.coli-Salmonella mixture at overnight incubation. 4. Among the diluted mixtures, 1:100, 1:1,000, and 1:10,000, survival rate of pathogenic enteric bacteria in the mixtures with E.coli showed least affected at the 1:1,000 dilution. 5. It was found that the antagonistic effect aganist groups of Salmonella-shigella was depending upon the groups of the genera.

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From the Sequence to Cell Modeling: Comprehensive Functional Genomics in Escherichia coli

  • Mori, Hirotada
    • BMB Reports
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    • v.37 no.1
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    • pp.83-92
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    • 2004
  • As a result of the enormous amount of information that has been collected with E. coli over the past half century (e.g. genome sequence, mutant phenotypes, metabolic and regulatory networks, etc.), we now have detailed knowledge about gene regulation, protein activity, several hundred enzyme reactions, metabolic pathways, macromolecular machines, and regulatory interactions for this model organism. However, understanding how all these processes interact to form a living cell will require further characterization, quantification, data integration, and mathematical modeling, systems biology. No organism can rival E. coli with respect to the amount of available basic information and experimental tractability for the technologies needed for this undertaking. A focused, systematic effort to understand the E. coli cell will accelerate the development of new post-genomic technologies, including both experimental and computational tools. It will also lead to new technologies that will be applicable to other organisms, from microbes to plants, animals, and humans. E. coli is not only the best studied free-living model organism, but is also an extensively used microbe for industrial applications, especially for the production of small molecules of interest. It is an excellent representative of Gram-negative commensal bacteria. E. coli may represent a perfect model organism for systems biology that is aimed at elucidating both its free-living and commensal life-styles, which should open the door to whole-cell modeling and simulation.

Detection of VTe-producing E coli using PCR method (PCR 기법 이용 VTe 분비 대장균 검출)

  • Yoon, Soon-seek;Park, Nam-yong;Lim, Jeong-taek
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.607-614
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    • 1996
  • Several methods for rapid and accurate detection of VTe-producing E coli were established. These methods contain beta-glucuronidase-secretion test, beta-haemolysis-production test in blood agar, verocytotoxicity test, and PCR. All of the VTe-producing strains made beta-haemolysis on 5% sheep blood agar. VTe-producing strains secreted beta-glucuronidase whereas 0157:H7 strains producing VTI or VTII did not secrete that enzyme. Verocytotoxicity test was established for rapid diagnosis. VTe detection was rapider in Vero cell suspension than Vero cell monolayer. In PCR, there was a positive result only in VTe-producing E coli, not in VTI or VTII-producing E coli. In this experiment, 165 strains of E coli were islated from feces or intestinal contents of post-weaning piglets showing nervous sign or diarrhea. And 20 strains of E coli that produced VTe were selected by verocytotoxicity test and PCR. According to these experiments, there was a direct correlation between verocytotoxicity test and PCR. And verocytotoxicity test is recommended as a routine diagnostic method and PCR does as a accurate diagnostic method to detect VTe-producing E coli.

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Molecular Cloning and Expression of Heat-stable Enterotoxin Gene from Swine Enterotoxigenic Escherichia coli (돼지에서 유래한 병원성 대장균의 내열성 장독소 생산유전자의 Cloning 및 발현)

  • 김교창;도대흥
    • Journal of Food Hygiene and Safety
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    • v.6 no.3
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    • pp.147-155
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    • 1991
  • Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin(ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxingeic E. coli from non pathogenic E. coli. The isolate of enterotoxigenlc E. coli was isolated from swine during 1989 year(from 5 to 10 month) in the Kyong-gi and Chung-Cheong provinces, and three strains(KM-4, KM-7 and KM-12) was selected from 189 isolates of ST producing E. coli. The detection of a ST produced of the isolated E. coli was performed by the infant mouse assay(IMA). This study was designed to know optimal conditions for the production of the ST and the molecular properties of plasmids of the enterotoxigenic E. coli. Amount of ST produced were the most at initial pH 8.5~9.0 of succinate salts medium culture. The cultural time of the same medium was accumulated the highest level of ST was at the 14 to 16 hours, and then stationary phase was at the 20 hours. From this experiment the KM-7 strain was selected among ST producing strains by IMA. Partial plasmid-curing experiment was done to select plasmid encoding for ST among other plasmids and then comparing the plasmid pattern of ST producing strain(KM-7) with those of other ST non-producing strains, it is found that ST gene exists on the about 80 Kbp plasmid. Each fragment of this plasmid digested with EcoRl was ligated to vector pBR 322 and transformed into E. coli K-12. A clone producing ST(eKT 53) was selected by IMA. The EcoRl digestion pattern of the isolated plasmid(pKD 37) from the ST producing clone it is indicated that the size of the inserted fragment in eKT 53 strain is 16 Kbp. The cultured supernatant of eKT 53 strain was positive result of ST production in IMA.

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Applicability Investigation of E.coli, RNA and DNA Bacteriophages for Possible Indicator Microorganisms Based on the Inactivation Effectiveness by UV (UV 불활성화 효과에 의거한 E.coli, RNA 및 DNA 박테리오파지의 대체 지표 미생물로서의 적용성 검토)

  • Kim, Il-Ho;Wahid, Marfiah AB;Tanaka, Hiroaki
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.11
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    • pp.1063-1068
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    • 2010
  • This study compared UV and UV/$H_2O_2$ inactivation of E.coli, a possible indicator microorganism for fecal contamination of water, and $Q{\ss}$ phage, an indicator for pathogenic viruses. UV inactivation of $Q{\ss}$, T4 and lambda phages in actual secondary effluent was investigated, too. As a result, similar inactivation efficiency between $Q{\ss}$ phage and E.coli was observed during UV treatment, while $Q{\ss}$ phage showed higher resistance to UV/$H_2O_2$ than E.coli. $Q{\ss}$ phage resistance to UV or UV/$H_2O_2$ does not reflect those of all pathogenic viruses. However, the result tells that the use of E.coli inactivation efficiency in evaluating microbiological safety of water could not always ensure the sufficient safety from pathogenic viruses. Meanwhile, $Q{\ss}$ phage showed less resistance to UV than T4 and lambda phages, indicating that the use of $Q{\ss}$ phage as an indicator virus may bring insufficient disinfection effectiveness by causing the introduction of lower UV dose than required. Consequently, it can be thought that T4 or lambda phages would be more desirable indicators in ensuring the sufficient disinfection effectiveness for various pathogenic viruses.