• Title/Summary/Keyword: Sucrose gradient

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Ozone Inactivation of Bacteriophage f2 (오존에 의한 bacteriophage f2의 살균작용(殺菌作用))

  • Kim, Chi-Kyung
    • Applied Microscopy
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    • v.11 no.1
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    • pp.29-38
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    • 1981
  • Bacteriophage f2 were treated with ozone at various concentrations for 20 minutes. The inactivation kinetics of f2 phage were examined during ozonation. In order to study the mode of action of ozone on the phage f2, absorption of the phage to the host pili was meassured by utilyzing radioactivity of tritium incorporated into the phage RNA. Sucrose density gradient analysis and electron microscopy were also used to prove the mechanism of ozone inactivation of the phage. Strucural proteins of the phage were broken by ozonation into many protein subunits. The extent of phage breakage was proportional to ozone concentration and reaction time. Percent decrease of the phage absorption to the host pili was coincident with the rate of ozone inactivation of the phage. Ozone inactivation of bacteriophage f2 was shown to be caused by the breakage of the structural protein and blockage of the phage absorption to the host pili.

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Killing Activity and Molecular Properties of Bacteriophage Sigma FA1 of Bacillus circulans (Bacteriophage Sigma FA1의 치사활성과 구조특성)

  • 김철호;김동수
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.553-560
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    • 1991
  • In the previous paper (10). a new temperate phage, Sigma FA1 had been isolated from B. circulans. Sigma FA1 had an icosahedral head with a diameter of about 70 nm, and a tail about 15 nm long, and beared a circularly permuted, linear duplex DNA. Signla FA1 killed sensitive cells by a single-hit process. Phage DNA injected into the cell immediately after infection was degraded slowly. Our results indicate that the killing action of Sigma FA1 is different from the phenomenon of abortive infection and suggest that the killing might be caused by a proteinaceous component of Sigma FA1.

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Genomic Variation and Toxin Specificity of Ustilago maydis Virus Isolated in Korea (한국에서 분리된 Ustilago maydis 바이러스의 유전자의 변이와 독소의 특이성)

  • Hee, Hwang-Seon;Yie, Se won
    • Korean Journal of Microbiology
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    • v.31 no.3
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    • pp.184-188
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    • 1993
  • Novel Ustilagomaydis strains, designated as SH1 to 14 containing new types of ds RNA segments, are identified from corn smut in Korea. Among 14 isolates, 7 isolates appear to posses virus particles and the other isolates may contain dsRNA as a plasmid form. The pattern of dsRNA is highly diverse form a typical P-type containing one or more of H, M, and L dsRNAs to the one containing one or move M dsRNAs. It is likely that the strains containing H dsRNA posses virus particles which were confirmed by sucrose density gradient followed with different range of specificity and the activity of the strain (SH14) is stronger than A4 toxin. The sensitivity of 14 isolates is also very diverse and two strains (SH10, SH11) appear tobe universal sensitve strains against 5 tested toxin samples.

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Sensitization Effects of Hyperthermia on Bleomycininduced DNA Strand Breaks and Replication Inhibition in CHO-$K_1$ Cells in Vitro

  • Kim, Chan-Gil;Kim, Hyeon-Suk;Park, Sang-Dai
    • Environmental Mutagens and Carcinogens
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    • v.15 no.2
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    • pp.88-93
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    • 1995
  • Effects of hyperthermia on the induction of DNA single strand breaks and replication inhibition were studied in bleomycin-treated CHO-K$_1$ cells by alkaline elution and alkaline sucrose gradient sedimentation. Bleomycin-induced DNA single strand breaks of DNA were dose-and time-dependently increased, and these strand breaks of DNA were gradually rejoined as post-incubation time passed. Treatment with hyperthermia alone did not affect the induction of DNA single strand breaks. However, pre-exposure of cells to hyperthermia followed by bleomycin treatment greatly increased the single strand breaks, and also reduced the rejoining processes of bleomycin-induced DNA single strand breaks. Bleomycin selectively inhibited the replicon initiation. The combined treatment with hyperthermia and bleomycin markedly potentiated the nonspecific inhibition of replication.

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Studies on Flacherie and Ina-flacherie Viruses of the Silkworm, Bombyx mori I. Purification of Viruses (가잠의 연화병바이러스에 관한 연구 I. 연화병바이러스의 정제)

  • 강석권
    • Journal of Sericultural and Entomological Science
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    • v.19 no.1
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    • pp.25-32
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    • 1977
  • The flacherie (FV) and Ina-flacherie (Ina-FV, named arbitarily) viruses of the silkworm, Bombyx mori was isolated from infectious larvae. Two types of infectious particles were purified by sucrose density gradient centrifugation. Some properties of the purified particles were investigated. Electron micrographs showed that FV and Ina-FV were spherical particles with diameters of 27nm and 20nm, respectively. The sedimentation coefficient of Ina-FV was 80S.

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Studies of Density-Fractionated Human Ervthrocvte Membranes (농도분배에 따라 분리한 사람 적혈구 막에 관한 연구)

  • 정종문
    • The Korean Journal of Zoology
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    • v.37 no.4
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    • pp.597-604
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    • 1994
  • Membranes obtained from the normal human RBC population were separated by continuous sucrose density gradient centrifugation and the density-fractionated membranes were then examined for changes in molecular markers. This study focuses on changes of (i) the membrane protein profile, (ii) differences in membrane-associsted enzyme activities, and (iii) the amount of autologous IgG bound. The following observations were made: (i) ratios for band 4. la over the sum of bands (4. la + 4.Ib) ranged from 0.58 to 0.79 for membranes of lowest density; (ii) significant changes in bound glyceraldehyde-3-phosphate dehydrogenase and acetvlcholinesterase activities were found; (iii) the amounts of autolosous IgG's attached to the red blood cells was highest in the membrane fraction of lowest density.

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Identification of novel $Ca^{2+}$ binding proteins in junctional sarcoplasmic reticulum of rabbit skeletal muscle

  • Jung, Dai-Hyun;Mo, Sang-Hyun;Kim, Do-Han
    • Proceedings of the Korean Biophysical Society Conference
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    • 2002.06b
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    • pp.56-56
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    • 2002
  • Muscle contraction and relaxation are regulated by the sarcoplasmic reticulum (SR)-mediated $Ca^{2+}$ release and $Ca^{2+}$ uptake. The SR functions are closely related with the proteins residing in the SR such as ryanodine receptor, $Ca^{2+}$-ATpase, calsequestrin, triadin and junctin. In an effort to further identify important functional SR proteins, experiments of sucrose-density gradient of SR fractionation, concanavalin A treatment, 2D gel electrophoresis, $^{45}$ Ca$^{2+}$ overlay, Strains-all staining, and peptide finger printing (PFP) were carried out.(omitted)d)

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Purification of Band 3 from the Human Erythrocyte Membrane and its Incorporation into Liposome (사람 적혈구막 Band 3의 정제 및 Liposome으로의 도입)

  • Kim, Jae-Ryong;Kim, Jung-Hye;Lee, Ki-Yung
    • Journal of Yeungnam Medical Science
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    • v.3 no.1
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    • pp.41-48
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    • 1986
  • Band 3, the predominent 95,000 dalton anion transport protein, is the major intrinsic glycoprotein of the human erythrocyte membrane. This anion carrier exists as a dimer and binds the cytoskeletons such as spectrin, ankyrin and actin. And the liposomes are vesicular structures which form spontaneouly upon hydration of phospholipids. These artificial lipid vesicles have been investigated as model of the biological membranes and as a mean of improving the delivery of nucleic acids, drugs, proteins and biological substances to specific target tissues and cells. In this study, we were purified Band 3 from the human erythrocyte membrane(ghost) was prepared by hemolysis of intact human erythrocyte with weak alkali-hypotonic solution. Band 6 was removed from ghost by extracting with solution of an ionic strength of 0.15. Band 3 and Band 4 were solubilized selectively by extracting Band 6-depleted ghosts with Triton X-100 under nondenaturing conditions. Band 3 was then purified from Triton X-100 extract treated with p-chloromercuribenzoate by sucrose density gradient ultracentrifugation. This purified Band 3 was incorporated into liposomes prepared by reverse-phase evaporation. Phosphatidyl L-serine and cholesterol(1 : 1 molar ratio) were dissolved in chloroform and then chloroform was removed by rotatory evaporation under reduced pressure. Band 3 solution without Triton X-100 was introduced into a mixture of lipids and diethylether. Diethylether was subsequently removed by evaporation. This purified Band 3 and its incorporation into liposomes were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

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The nature of non-specific inhibitor and natural agglutinin for goose erythrocytes in chicken serum (계혈청(鷄血淸)에 들어 있는 뇌염(腦炎)바이러스에 대한 비특이물질 및 거위 혈구(血球)에 대한 응집소의 성상(性狀)에 관한 연구(硏究))

  • CHUNG, Y.S.
    • Korean Journal of Veterinary Research
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    • v.11 no.2
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    • pp.123-136
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    • 1971
  • (1) The non-specific inhibitors (NSI) in normal chicken sera were active against all the tested group A and group B arboviruses, but the group B arbovirus were more sensitive than group A arboviruses. (2) The titres of the NSI were distributed nearly uniformly among chickens from seven different age groups to group A arboviruses. In contrast, the NSI titres to group A arboviruses were found to increase with age. (3) No significant difference could be demonstrated between acetone-ether extraction and kaolin adsorption for removal of the NSI in normal chicken sera. (4) After heating, the NSI titres in chicken sera were increased for both group A and group B arboviruses. (5) After heating the sera at $80^{\circ}C$ and $100^{\circ}C$, kaolin adsorption was less efficient for removing the NSI than it, was in unheated serum. Acetone-ether extraction of the NSI was unimpaired after heating at $80^{\circ}C$ but was less efficient after heating at $100^{\circ}C$. (6) The NSI activity was found mainly in the first peak (IgM) and diffused to a part of second peak (IgG) by fractionation of chicken serum by gel filtration through Sephadex G200. After zonal centrifugation of chicken serum in a linear ten to 40 percent sucrose gradient all of the NSI activities were found on the top of the centrifugal tubes. These properties of large molecular size and low density indicated that the NSI in chicken serum were probably lipoproteins. (7) The natural agglutinins for goose erythrocytes in chicken sera were partially destroyed by acetone-ether extraction but not by kaolin adsorption, and were efficiently adsorbed with ten percent goose erythrocytes. No difference of the NA titre was demonstrated with diluents of different pH. (8) The NA in chicken serum was found to possess the properties of IgM by gel filtration through Sephadex G200 and zonal centrifugation in linear ten to 40 percent sucrose gradient.

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Isolation of Cymbidium mild mosaic virus (Cymbidium mild mosaic virus의 분리동정)

  • Chang M. U.;Doi Y.;Yora K.
    • Korean journal of applied entomology
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    • v.17 no.3 s.36
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    • pp.131-138
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    • 1978
  • A virus named Cymbidium mild mosaic virus(Cy MMV), was mechanically transmitted to Chenopodium amaranticolor from the leaves of Cymbidium with mild mosaic symptoms. The virus was cultured in C. amaranticolor, in which it produced local chlorotic and ring spots, followed by systemic vein clearing with distortion. CyMMV infected 7 out of 35 species of plants. In C. amaranticolor juice infectivity was lost by heating at $90^{\circ}C$ for 10 miuntes, and by aging at$20^{\circ}C$ for 60 days, and by diluting at $10^{-6}$ when bioassayed on C. amaranticolor. CyMMV was not transmitted by Myzus persicae. The virus was purified after clarification of homogenized C. amaranticolor leaf tissues with chloroform, by differential centrifugation followed by sucrose density gradient centrifugation. Electron microscopic examination of purified preparation showed spherical particles of 28nm in diameter. The UV absorption spectrum of purified preparation was typical of u nucleoprotein (max. at 261nm. min. at 243nm), and showed 260/280=1.72 and max/min=1.26. The value of the sedimentation coefficient of the virus was S20.w=126. In gel-diffusion tests, CyMMV antiserum reacted with CarMV, but not with any of four other viruses (BBWV, CRSV, CMV, TBRV) having similar particles and properties in vitro. In ultra-thin sections of CyMMV infected tissues, a large number of virus particles were found in the cytoplasm of mesophyll cells and in xylem vessels.

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