• Title/Summary/Keyword: restriction endonuclease analysis

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Comparison in Restriction Profile Analysis of Vibrio furnissi, Vibrio fluvialis, and Vibrio parahaemolyticus Bacteriophage from Sea Product

  • Kim, Young-Hee
    • Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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    • v.1 no.2
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    • pp.99-103
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    • 1997
  • The bacteriophages lytic for Vibrio furnissi, Vibrio fluvialis and Vibrio parahaemolyticus were isolated from fish gills and shellfish. Nucleic acid of bacteriophages was prepared and restriction endonuclease profile was compared. All isolates contained deoxyribonucleic acid. V. furnissi bacteriophage from fish gills showed 2 bands with Bgl II, 1 with Pst, 3 with Hind III, I with Bam HI and 2 with EcoR 1. V fluvialis phage represented 7 fragments with Bgl II, 1 with Pst, 4 with Hind III, and 2 with EcoR 1. V. parahamolyticus produced 13 sites with Hind III and 4 sites with EcoR 1. The fragment types were varied depending on the phage isolation. All three phages were digested with Hind III and EcoR I with different sizes. V. furnissi phage were digested with 5 different restriction enzymes.

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Molecular epidemiologic analysis of pathogenic Escherichia coli isolated from poultry in Korea (국내 가금 유래 병원성 대장균의 분자역학적 분석)

  • Sung, Myung-Suk;Kim, Jin-Hyun;Kim, Ki-Seuk
    • Korean Journal of Veterinary Research
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    • v.50 no.3
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    • pp.239-246
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    • 2010
  • Among 203 avian pathogenic Escherichia coli (APEC) isolated from poultry with colibacillosis in korea, 14 isolates were selected from total 68 isolates transferred R plasmid and classified into 5 groups on the basis of antimicrobial minimal inhibitory concentration (MIC) pattern, farm source and O serotype. An association between clonal origin and R plasmid of them was investigated by R plasmid profile, restriction endonuclease analysis and pulsed-field gel electrophoresis (PFGE). The strains that showed the same or very similar antimicrobial MIC pattern, but different farm source and O serotype, revealed different PFGE pattern, which seemed to be different clonal origin. And the strains that showed the same MIC pattern and O serotype, revealed different PFGE pattern, seemed to be originated from different clone. Also the strains showing the same MIC pattern and farm source, but different O serotype, revealed to be different clonal origin. The strains that showed the same or similar MIC pattern, farm source, and O serotype, revealed identical or similar PFGE pattern, which seemed to belong to be one clone. Meanwhile, horizontal transfer of R plasmid seems to be common in APEC with regardless of O serotype and clone of the strains. These results indicate that rapid and accurate epidemiological survey of APEC can be possible by the combination of O serotyping, plasmid profiling and PFGE analysis following the classification of them into groups of antimicrobial drug resistance pattern.

Molecular Characteristics of R Plasmids in Shigella (Shigella R Plasmid의 분자적 특성)

  • Lee, Yoo-Chul;Seol, Sung-Yong;Cho, Dong-Taek;Chun, Do-Ki
    • 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.

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A Profile of Naturally Occurring Plasmids from Selected Strains of Vibrios

  • Younghee Kim
    • Journal of Environmental Science International
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    • v.1 no.2
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    • pp.93.2-97
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    • 1992
  • The naturally occurring plasmids of Vibrio species have been isolated in part to investigate their genetic traits. Among six different Vibrio species tested, Vibrio anguillarum, Vibrio fluvialis, Vibrio vulnificus, Vibrio mimicus and Vibrio furnissi did not show any presence of plasmid. One environmental isolate of Vibrio pnrahemolyticus harboring plasmid was observed. The isolated plasmid was 8.7 kb by analysis with restriction endonuclease digestion. No common feature was shown relationships between the presence of plasmid and resistance against commonly used antibiotic compounds from the tested Vibrios. Key words . plasmid, Vibrio anguillarum, Vibrio fluvialis, Vibrio uulnincus, Vibrio mimicus, Vibrio furnissi, Vibrio parahemolyticus.

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A Profile of Naturally Occurring Plasmids from Selected Strains of Vibrios

  • Kim, Young-Hee
    • Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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    • v.1 no.2
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    • pp.93-97
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    • 1997
  • The naturally occurring plasmids of Vibrio species have been isolated in part to investigate their genetic traits. Among six different Vibrio species tested, Vibrio anguillarum, Vibrio fluvialis, Vibrio vulnficus, Vibrio mimicus and Vibrio furnissi did not show any presence of plasmid. One environmental isolate of Vibrio parahaemolyticus harboring plasmid was observed. The isolated plasmid was 8.7 kb by analysis with restriction endonuclease digestion. No common feature was shown relationships between the presence of plasmid and resistance against commonly used antibiotic compounds from the tested Vibrios.

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The Genetic Diversity of Bacterial Communities in the Groundwater (지하수 세균 군집의 유전적 다양성)

  • 김여원;민병례;최영길
    • Korean Journal of Environmental Biology
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    • v.18 no.1
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    • pp.53-61
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    • 2000
  • In order to characterize the genetic diversity of bacterial community in groundwater, samples were collected from used for drinking water and polluted with heavy metal wastewater in Seoul city and natural cave of Kangwondo. The DNA was amplified with 165 rDNA-based primers by use of the PCR, and then analysed ARDRA (amplified ribosomal DNA restriction analysis). Restriction endonuclease analysis patterns of amplified 165 rDNA in drinking water and wastewater relatively showed high genetic diversity in situ and drinking groundwater. The number of DNA fragments varied with in situ and drinking water. This method of ARDRA of bacterial communities in groundwater could be used for a quick assessment of genotypic changes between different locations reflecting different environmental conditions and the diversity reflected pollution of groundwater (natural cave water>drinking water>waste water, as in order of grade). [Genetic diversity, Groundwater, 165 rDNA, PCR, ARDRA].

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Morphology and Biochemcial Characteristics of a Nuclear Polyhedrosis Virus Isolated from the Oriental Tobacco Budworm, Helicoverpa assulta (Guenee) (담배나방 핵다각체병바이러스의 형태 및 생화학적 특성)

  • 진병래;박현우;우수동;김우진;김우진;박범석;강석권
    • Korean journal of applied entomology
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    • v.34 no.3
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    • pp.218-223
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    • 1995
  • A nuclear polyhedrosis virus isolated from the oriental tobacco budworm larvae, Helicoverpa assulta (Guenee) was characterized by electron microscopy, SDS-PAGE, restriction endonuclease analysis and cross infectivity. The shape of a polyhedron was $1.0\mu\textrm{m}$ in average with icosahdral outline, and the virus particle was $65nm\times300nm$ in average with rod-shape. The nuclear polyhedrosis virus was contained a single nucleocapsid within a viral envelope embedded in a polyhedron. The polyhedral protein was composed of a single polypeptide with a M.W. of 31 Kd. The genome size of the virus by restriction endonuclease analysis was about 120 Kb. Among several nuclear polyhedrosis viruses, the nuclear polyhedrosis virus from Helicoverpa assulta (HaNPV) and Autographa california nuclear polyhedrosis virus (AcNPV) were infected the oriental tobacco budworm larvae.

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Biochemical Characteristics of the Nuclear Polyhedrosis Viruses of the Fall Webworm, Hyphantria cunea, and the Silkworm, Bombyx mori (누에와 흰불나방 핵다각체병바이러스의 생화학적 특성)

  • 김현욱;박범석;진병래;임대준;강석권
    • Korean journal of applied entomology
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    • v.28 no.3
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    • pp.105-112
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    • 1989
  • The nuclear polyhedrosis viruses of Bombyx mori (BmNPV) and Hyphantria cunea (HcNPV) were characterized by electron microscopic observation, SDS-PAGE of polyhedral and virion proteins, and restriction endonuclease analysis of viral DNAs. Polyhedra of BmNPV were octadecahedral in shape with the diameter of $3 \mu\textrm{m}$, whereas those of HcNPV showed irregular appearances having the diameter of $1.5~2\mu\textrm{m}$. Under alkaline protease inactivated condition, polyhedral proteins of two NPVs were resolved into a major polypeptide, 30~31 KD, and several minor polypeptides by SDS-PAGE. Examination of virion proteins by silver staining after SDS-PAGE showed that BmNPV was composed of 47 polypeptides with M.W. range of 9.6~112 KD and HcNPV was composed of 48 polypeptides with M.W. range of 9.4~111 KD. The approximate genome size of two NPVs were determined by restriction endonuclease analysis: BmNPV and HcNPV were 114.6 Kb, respectively.

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Molecular cloning and restriction endonuclease mapping of homoserine dehydrogenase gene (HOM6) in yeast saccharomyces cerevisiae (Aspartate계 아미노산 대사 효모 유전자 HOM6의 cloning 및 구조분석)

  • 김응기;이호주
    • Korean Journal of Microbiology
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    • v.24 no.4
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    • pp.357-363
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    • 1986
  • Synthesis of threonine and methionine in yeast, Saccharomyces cerevisiae shares a common pathway from aspartate via homoserine. HOM6 gene encodes homoserine dehydrogenase (HSDH) which catalyzes the inter-conversion of beta-aspartate semialdehyde and homoserine. The level of HSDH is under methionine specific control. A recombinant plasmid (pEK1: 13.3kb), containing HOM6 gene, has been isolated and cloned into E. coli by complenemtary transformation of a homoserine auxotrophic yeast strain M-20-20D (hom6, trp1, ura3) to a prototrophic M20-20D/pEK1, using a library of yeast genomic DNA fragments in a yeast centromeric plasmid, YCp50(8.0kb). Isolation of HOM6has been primarily confirmed by retransformation of the original yeast strain M20-20D, using the recombinant plasmid DNA which was extracted from M20-20D/pEK1 and subsequently amplified in E. coli. Eleven cleavage sites in the insery (5.3kb) have been localized through fragment analysis for 8 restriction endonucleases; Bgl II(2 site), Bgl II(1), Cla I(3), Eco RI(1), Hind III(2), Kpn I (1), Pvu II(1) and Xho I(1).

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Characterization of Plasmid Encoding a Heat-labile Enterotoxin Originated from an Human Escherichia coli Strain O15: H11. (사람에서 유래한 장독성 대장균의 이열성장독소와 관련된 Plasmid 특성)

  • Rhyu, Mun-Gan;Kim, Gum-Ryong;Chang, Woo-Hyun
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.47-52
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    • 1986
  • A heat-labile enterotoxin and no heat-stable enterotoxin producing($LT^+ST^-$) plasmid (110 kilobases in size) was isolated from an enterotoxigenic Escherichia coli of human strain O15:H11 and used for analysis of the $LT^+$ deoxyrionucleic acid region using recombinant DNA technology. A DNA segment containing the $LT^+$ DNA region which was one restriction endonuclease BamHl fragment(6.2 kb in size) was joind to a small multicopy plasmid, pUC9. E. coli K-12 strain, JM103 harboring the chimeric plasmid produced greater amounts of LT than did the enterotoxigenic E. coli O15:H11 strain. The BamHl fragment was further digested with various restriction endonucleases and contained no HindIll restriction site which is an essential in $LT^+ST^+$ plasmid. The detailed DNA sequencing of this $LT^+ST^-$ plasmid is required.

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