• Title/Summary/Keyword: ethidium bromide

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Effect of Ethidium Bromide on Anomalous Absorbance-Temperature Profile, of Calf Thymus DNA, induced by Spermine (송아지 胸線 DNA의 Spermine에 依하여 誘發되는 異例的 吸光度-溫度 樣相에 미치는 Ethidium Bromide의 影響)

  • Chan Yong Lee;Thong Sung Ko
    • Journal of the Korean Chemical Society
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    • v.29 no.3
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    • pp.271-276
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    • 1985
  • An anomalous absorbance-temperature profile of calf thymus DNA, having a trough preceding the initiation of the melting, occurs at the spermine concentration, where the DNA collapses into a compact structure. The cooperativity, enthalpy, and the midpoint of the phase transition to the trough region are more sensitive to ethidium bromide than those of the Tm region. As the concentration of ethidium bromide added is increased, the peak size of the trough is decreased, while the Tm remains essentially constant.

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Elimination of R-Plasmid in Streptomyces bobili (YS-40) by Ethldium Bromide (Ethidium Bromide에 의한 Streptomyces bobili(YS-40)의 R-Plasmid 제거)

  • 김상달;도재호
    • Microbiology and Biotechnology Letters
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    • v.10 no.4
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    • pp.289-295
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    • 1982
  • Streptomyces bobili (YS-40) isolated from soil was tested that it had drug resistance against penicillin, cephalosporin series antibiotics and other antibiotics in the previous paper. The treatment of Streptomyces bobili, (YS-40) with ethidium bromide (EtBr), acriflavine and sodium dodecyl sulfate. (SDS) resulted in the elimination of R-plasmid from the host strain. Minimum growth inhibitory concentrations (MIC) of Hg, Ag, penicillin-G, ampicillin, chloramphenicol, oxytetracycline, streptomycin and kanamycin were found to be 15, 10, > 3, 000, > 100, > 1, 000, > 100, < 5 and < 5$\mu\textrm{g}$/$m\ell$ respectively. Among the curing agents, EtBr was proved to be the most powerful compound for the elimination of R-plasmid in the strain and the elimination rate with EtBr(10$\mu\textrm{g}$/$m\ell$) was about 98%. Optimal pH to. the elimination of R-plasmid was pH 7.0 and the R-plasmid in the cells incubated for 24 hrs was proved to be eliminated most effectively. Aerial mass color, soluble pigment formation and reverse side color were reported to be often the plasmid associated characteristics of the R-plasmid bearing bacteria. But these characteristics of the uncured and cured Streptomyces bobili, (YS-40) showed no changes in the most of the pigment formation media tested in this work.

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Curing Action of Antibiotic Resistant Factor in Bacillus Subtilis (Bacillus subtilis의 항생물질 내성에 대한 Curing작용)

  • Hong, Yong-Ki;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.13 no.2
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    • pp.103-107
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    • 1985
  • A variety of plasmid curing agents such as sodium dodecyl sulfate, acriflavine, ethidium bromide, and mitomycin-C were used to cure Bacillus subtilis cells of streptomycin resistant factor. The drug susceptibility was increased by 0.1% sodim dodecyl sulfate at stationary growth phase. The curing frequency was obtained highly at 4 $\mu\textrm{g}$/$m\ell$ of acriflavine, 10 $\mu\textrm{g}$/$m\ell$ of ethidium bromide, and 200 $\mu\textrm{g}$/$m\ell$ of mitomycin-C. respectively. Curing action occurred competitively for the streptomycin and terramycin resistant factors in B. subtilis.

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Improved Fluorometric Assay Method for Ribonuclease Activity

  • Lee, Jong-Soo;Choi, Jong-Soo
    • BMB Reports
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    • v.30 no.4
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    • pp.258-261
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    • 1997
  • A simple quantitative assay method for ribonuclease activity has been developed. This method is based on the decrease of fluorescence intensity emitted by the ethidium bromide bound to RNA due to the degradation of RNA by ribonuclease. The substrate RNA was reacted with ribonuclease A and the fluorescence intensity was measured after the addition of ethidium bromide. The intensity difference was calculated using a blank reaction mixture containing no RNase. Whole cellular RNA substrate produced a significant error and was not suitable for this assay method possibly because of local microheterogeniety caused by high molecular weight rRNA. but satisfying results were obtained with tRNA substrate. The intensity difference increased linearly by raising enzyme concentration up to $2{\times}10^{-4}$ Kunitz Units of ribonuclease A. More refined and reliable results were obtained by use of initial reaction velocities which were calculated from the plots of intensity difference vs time. A linear relationship between initial velocities and enzyme concentrations was observed up to 0.01 Kunitz Units of enzyme.

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Rapid Isolation of Mitochondrial DNA-Depleted Mammalian Cells by Ethidium Bromide and Dideoxycytidine Treatments

  • Yoon, Young Geol;Oh, Yoo Jin;Yoo, Young Hyun
    • Journal of Applied Biological Chemistry
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    • v.57 no.3
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    • pp.259-265
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    • 2014
  • Mitochondrial DNA (mtDNA)-depleted (${\rho}^0$) cells are often used as mtDNA recipients to study the interaction between the nucleus and mitochondria in mammalian cells. Therefore, it is crucial to obtain mtDNA-depleted cells with many different nuclear backgrounds for the study. Here, we demonstrate a rapid and reliable method to isolate mammalian mtDNA-depleted cells involving treatment with the antimitochondrial agents ethidium bromide (EtBr) and 2',3'-dideoxycytidine (ddC). After a short exposure to EtBr or ddC, followed by rapid clonal isolation, we were able to generate viable mtDNA-depleted cells from mouse and human cells and were able to successfully repopulate them with exogenous mitochondria from platelets isolated from mouse and human blood samples. These mtDNA-depleted cells can be used to characterize the nuclear mitochondrial interactions and to study mtDNA-associated defects in mammalian cells. Our method of isolating mtDNA-depleted cells is practical and applicable to a variety of cell types.