• Title/Summary/Keyword: Agarose gel

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Apoptotic Cell Death of Mouse Splenocytes by Polychlorinated Biphenyls and Its Prevention by Serum

  • Yoo, Byung-Sun;Kim, Hwan-Mook
    • Toxicological Research
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    • v.13 no.3
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    • pp.187-191
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    • 1997
  • Cell death induced by polychlorinated biphenyls (PCBs), environmental toxicant, was investigated in mouse splenocytes. The fragmentation of intact DNA, a parameter of apoptotic cell death, was evaluated qualitatively by agarose gel electrophoresis analysis and quantitatively by diphenylamine reaction method. PCBs induced apoptotic cell death of splenocytes in a dose- and time- dependent manner. The effect of serum on the apoptotic cell death induced by PCBs was also investigated. The DNA fragmentation induced by PCB treatment in serum-free medium was clearly inhibited by an addition of serum to the culture medium. The decrease of DNA fragmentation due to serum addition was accompanied with the increase of cell viability.

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Biodegradative Characteristics of Benzoate and m-Toluate by Pseudomonas sp. (Pseudomonas sp.에 의한 Benzoate와 m-Toluate 의 분해특성)

  • 정준영;김교창
    • Journal of Food Hygiene and Safety
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    • v.9 no.4
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    • pp.205-211
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    • 1994
  • From 120 soil and activated sludge, the strains able to grow on benzoate and m-Toluate have been isolated after selective enrichment which were later identified as Psudomonas sp. according to its morphological and biochemical characteristics. Ben-2 strain contained two plasmid DNA having about 120 Kb and below 2.0 Kb by agarose gel electrophoresis. Form the comparative investigation of catechol 1,2-oxygenase and catechol 2,3-oxygenase activities in Ben-2 strain and its cured strain, Ben-2 strain has both of these two enzymes while cured strain has catechol 1,2-oxygenase only.

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Effects of Neutral, Cationic, and Anionic Chromium Ascorbate Complexes on Isolated Human Mitochondrial and Genomic DNA

  • Ay, Ahmet Nedim;Zumreoglu-Karan, Birgul;Oner, Reyhan;Unaleroglu, Canan;Oner, Cihan
    • BMB Reports
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    • v.36 no.4
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    • pp.403-408
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    • 2003
  • The relative activities of neutral, cationic, and anionic chromium ascorbate complexes toward isolated human mitochondrial and genomic DNA were investigated at physiologically relevant conditions by agarose gel electrophoresis. A direct relationship between the charge of the Cr(III) species and their DNA-damaging properties was found. The cationic species were found to be fully capable of DNA-cleavage, even in short incubation periods. Incubations were also performed in the presence of amino acids. No apparent effect was observed under the applied experimental conditions to facilitate or prevent damage through the ternary amino acid-Cr-DNA adduct formation or binary chromium-amino acid complex formation.

Tetra Primer ARMS PCR Optimization to Detect Single Nucleotide Polymorphisms of the CYP2E1 Gene

  • Suhda, Saihas;Paramita, Dewi Kartikawati;Fachiroh, Jajah
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3065-3069
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    • 2016
  • Single nucleotide polymorphism (SNP) detection has been used extensively for genetic association studies of diseases including cancer. For mass, yet accurate and more economic SNP detection we have optimized tetra primer amplification refractory mutation system polymerase chain reaction (ARMS PCR) to detect three SNPs in the cytochrome P450 2E1 (CYP2E1) gene locus; i.e. rs3813865, rs2070672 and rs3813867. The optimization system strategies used were (1) designing inner and outer primers; (2) determining of their optimum primer concentration ratios; and (3) determining of the optimum PCR annealing temperature. The tetra primer ARMS PCR result could be directly observed using agarose gel electrophoresis. The method succesfully determined three SNPs in CYP2E1 locus, the results being consistent with validation using DNA sequencing and restriction fragment length polymorphisms (RFLP).

Isolation and Characterization of Marinobacter sp. KS-1, which Produces a Chondroitin Sulfate-like Mucopolysaccharide

  • Lee, Dae-Sung;Kim, Kyung-Suk;Lee, Myung-Suk;Kim, Young-Mog
    • Fisheries and Aquatic Sciences
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    • v.13 no.3
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    • pp.210-215
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    • 2010
  • In an effort to isolate a bacterium producing chondroitin sulfate (CS), a marine bacterium, KS-1, which produces mucopolysaccharides, was isolated from seawater and identified as Marinobacter sp. based on analyses of its morphological and biochemical traits and 16S rDNA sequence. Agarose-gel electrophoresis showed that the KS-1 strain produces a CS-like mucopolysaccharide. Structural analysis using Fourier transform infrared spectroscopy revealed that the structure of the CS-like mucopolysaccharide produced by Marinobacter sp. KS-1 is similar to that of dermatan sulfate (CS B). However, the molecular mass of the CS-like mucopolysaccharide is higher than that of standard chondroitin sulfates. Considering the above results, we conclude that the Marinobacter sp. KS-1 produces a CS-like mucopolysaccharide that differs somewhat from CS B in molecular mass.

The Role of Lipid Peroxidation and Glutathione on the Glycochenodeoxycholic Acid-Induced Cell Death in Primary Cultured Rat Hepatocytes

  • Chu, Sang-Hui;Park, Wol-Mi;Lee, Kyung-Eun;Pae, Young-Sook
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.2
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    • pp.121-127
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    • 2000
  • Intracellular accumulation of bile acids in the hepatocytes during cholestasis is thought to be pathogenic in cholestatic liver diseases. The objective of this study was to determine the role of lipid peroxidation and glutathione on the bile acid-induced hepatic cell death mechanism in primary cultured rat hepatocytes. To induce hepatic cell death, we incubated primary cultured rat hepatocytes with glycochenodeoxycholic acid $(GCDC;\;0{\sim}400\;{\mu}M)$ for 3 hours. In electron microscopic examination and agarose gel electrophoresis, low concentration of GCDC treatment mainly induced apoptotic feature. Whereas $400\;{\mu}M$ GCDC treated cells demonstrated both apoptosis and necrosis. Lipid peroxidation was increased dose-dependently in GCDC treated hepatocyte. And this was also accompanied by decreased glutathione. Therefore, oxygen free radical damage may play a partial role in GCDC-induced hepatic cell death.

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Identification of Fungus from Dog and Diagnosis using PCR (개에서 분리한 진균의 동정과 PCR을 이용한 진단)

  • 장화석;문영찬;이상원;김휘율;김태종
    • Journal of Veterinary Clinics
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    • v.21 no.1
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    • pp.35-44
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    • 2004
  • This study was performed to examine the mycological features of canine skin. A total of 50 dogs with skin lesions were examined for dermatology from October, 2000 to April, 2001. The isolation rates of dermatophytes, yeast, filamentous fungi and superficial fungi were 36.4%, 13.5%, 35.3% and 13.6%. The dermatophytes isolated in dogs were Microsporum canins and Trichophyton mentagrophytes were 75% and 25%. The yeast and superficial fungi isolated in dogs were Candida albicans, Rhodntorula minnata, Candida ceferrii and Malassezia spp. were 16.7%. 16.7%, 16.7% and 50%. The filamentous fungi by Aspergillus funigatus, Aspergillus niger, Penicillum spp., Alternaria spp. were 12.5%, 12.5%, 50%, and 25%. In determine if polymerase chain reaction (PCR) could be applied for diagnosis of dermatophytes, yeast and filamentous fungi, control and clinical samples were tested. The size of specific PCR product in agarose gel was 340 bp for dermatophytes and 210 bp for yeast and filamentous fungi, respectively.

Effect Of Taraxacum coreanum Nakai on Low Density Lipoprotein Oxidation (흰민들레의 동맥경화 유발인자인 저밀도 지질단백질 산화에 미치는 영향)

  • Jeon, Cheol-Min;Yang, Ki-Sook
    • Korean Journal of Pharmacognosy
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    • v.27 no.3
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    • pp.267-273
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    • 1996
  • The methanol extract of Taraxacum coreanum Nakai (Compositae) was examined on the in vitro oxidation of human low density lipoprotein (LDL). It is well known that LDL oxidation induced artherosclerosis, if we can protect LDL oxidation process, excess plasma lipoprotein accumulation into the arterial lesion prone areas can be blocked. The methanol extract was treated with oxidized LDL which was incubated with $16\;{\mu}M$ $Cu^{2+}$ for metal catalyzed oxidation and TBA value, mobility on agarose gel and formation of conjugated diene and change of vitamin E were determined for the evaluation. The extract showed antioxidative effect at concentration $200\;{\mu}g/ml$ on LDL oxidation.

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Restriction pattern of the nucleic acid of Synechococcus sp. cyanophage

  • Park, Jong-Geun;Kim, Min;Choi, Yong-Keel;Yoon, Sung-Nyo
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.1-6
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    • 1996
  • The nucleic acid of Synechococcus sp. cyanophage was identified as double-stranded DNA by the result of digestion with enzymes such as exonucleases, DNase, and S1 nuclease, and by acridine orange staining. The cyanophage DNA was cleaved with several restriciton ehdonucleases such as ApaI, BamHI, Bg/II, HaeIII, Eco RI, HindIII, PstI, AND aPAI gave the clearest sets of bands on agarose gels and the fragment numbers for each were 12, 20, 29, 20, and 7, respectively. The sums of the size from Bam HI and PstI digestions were estimated approximately 227$\pm$4 kb, which are in agreement with the result of the pulsed field gel electrphoresis. This virus is thought to have the largest genosome among those of known cyanophages, which corresponds to the largest haed ot 90 nm when compared with the head sizes of cyanophages discovered since 1963.

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Genetic Characteristics of Arsenic Compounds-Resistant Bacteria Isolated from Stream Water (하천에서 분리한 비소 내성세균의 유전적 특성)

  • 정미경;이호자
    • Korean Journal of Microbiology
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    • v.29 no.1
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    • pp.63-68
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    • 1991
  • Several arsenic compound-resistant bacteria were isolated from Jung-Rang stream. The isolates, D-3, D-12, and D-14 were characterized phenotypically and genetically, and identified as Serratia liquefaciens, Klebsiella oxytoca, and Klebsiella pneumoniae, respectively. A plasmid of 67kb was found in Klebsiella oxytoca D-12 and designated as pMH12. Transfer of this plasmid from D-12 to E. coli HB101 was occurred, and the resulting transconjugant strains expressed the same level of heavy metal resistance as the donor strain. The physical presence of this plasmid in transconjugant was detected with agarose gel electrophoresis. Arsenite-sensitive derivatives of isolate D-12 were obtained with Mitomycin C treatment which cured pMH12. Antibiotics and heavy metal resistances were also examined to be used as a proper marker for the isolates in gene cloning. Isolate D-12 has resistance to several heavy metal ions such as $Cd^{2+}$ , $Zn^{2+}$ and $Hg^{ 2+}$ Also, all the other arsenite resistant isolates showed resistance to several heavy metal ions and antibiotics.

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