• Title/Summary/Keyword: Sucrose gradient

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Isolation of Cryptosporidium parvum oocysts from fecal samples - The combination of ether extraction and discontinuous sucrose gradients (분변에 함유된 Cryptosporidium parvum 오오시스트의 순수분리 -Ether extraction과 discontinuous sucrose gradients의 병용)

  • 위성환;이정길
    • Parasites, Hosts and Diseases
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    • v.32 no.1
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    • pp.7-12
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    • 1994
  • A calf and 50 mice were infected with Cryptosporidium parvum and their fecal materials were collected and treated 10 ether extraction (EE), followed by discontinuous sucrose gradients (DSG) method. EE method was to remove some of fat or lipid from feces. Sediments were washed by centrifugation ($1,500{\;}{\times}{\;}g$ for 10 min., 3 times) In phosphate-buffered saline and then these washed sediments were sleeved sequentially through stainless steel screens with a final mesh of 250 ($61{\;}{\mu\textrm{m}}$ porosity) to remove other debris. After sieving, the materials were suspended in 2.5% potassium dlchromate solution. Oocysts were counted by using a hemocytometer and the recovery rate of pure oocysts was calculated on the basis of the count. Following centrifugation ($1,500{\;}{\times}{\;}g$ for 30 min.) by DSG method, most oocysts were recovered at the interface between a gravity of 1.103 and 1.064. The recovery rates of pure oocysts from the fecal suspension of the calf ($3.8{\;}{\times}{\;}10^7/ml$) and the mouse ($3.2{\;}{\times}{\;}10^6/ml$) treated with EE method were 81.6% and 51.6%, respectively. It is suggested that the recovery rate was dependent on the number of oocysts In each suspension treated with EE method. To get the 50% recovery rate, there must be more than $2{\;}{\times}{\;}10^6$ oocysts per ml of the fecal suspension treated with EE method. By the combination of the two methods it was possible to isolate C. parvum oocysts from normal feces of the calf and mouse as well as from dlarrhelc feces.

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Histidylleucine Hydrolyzing Enzyme : A Dipeptidase From Hog Lung (Dipeptidase로서의 Histidylleucine 가수분해 효소)

  • Hyun-Jae Lee;Jung-Keun Park
    • Journal of the Korean Chemical Society
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    • v.19 no.5
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    • pp.375-380
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    • 1975
  • An enzyme capable of hydrolyzing histidylleucine was purified 50 fold from hog lung. The final preparation hydrolyzed $1.6{\mu}moles$ of histidylleucime per minute per mg of protein. The $K_m$ of the enzyme for the enzyme was found to be $2{\times}10^{-4}M$. The enzyme was required a number of free dipeptides for the substrate specificity, and was inhibited by EDTA and 1,10-phenan-throline. The molecular weight of the enzyme was estimated to be 80,000 daltons from sucrose density gradient sedimentation analysis. The corrected $s_{20,w}$ value was 5.3 S.

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Isolation and Identification of Cyanophage from Eutrophic Water (부영양화 수역에서의 Cyanophage 의 분리와 동정)

  • Kim, Min;Choi, Yong-Keel
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.524-527
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    • 1992
  • Synechococcus sp. cyanophage was isolated from Baekwoon reservoir located in KyonggiDo. The cyanophage was purified by employing ultrafiltration. differential centrifugation. and sucrose density gradient centrifugation. Electron microscopic observation indicated that the sizes of its isometric head and contractile tail are 89 nm and] II nm. respectively. which means that the isolated cyanophage is included in the group. Myoviridae. The cyanophage maintained the stability of more than 50 percent from $20^{\circ}C$ to $40^{\circ}C$ and from pH 5 to 8. and had the maximal infectivity at $30^{\circ}C$ and pH 9 implying its ecological significance.

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Esterase Isozyme of Mycelium of Pyricularia oryzae under Various Cultural Conditions (배양조건에 따른 도열병균(Pyricularia oryzae) 균사체의 Esterase 동위효소)

  • Park Sang Ho;Park Won Mok;Kim Seong Hoe;Lee Eun Jong
    • Korean Journal Plant Pathology
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    • v.3 no.3
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    • pp.168-173
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    • 1987
  • The constancy of the patterns of esterase isozymes in the mycelium of Pyricularia oryzae grown under various cultural conditions was observed by $10\~25\%$ polycrylamide gradient slab gel. The position of the isozyme was not affected by cultural age for 24 days. However the intensity of the band was affected. The band patterns were not affected by the carbon sources (glucose, maltose, fructose, sucrose or starch), single spore isolates, successive transfer culture and regenerated isolates from protoplast cultures.

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Bacteriophage-like Particles Induced by Mitomycin C in Bacillus circulans F-2 (Mitomycin C에 의해 유도되는 Bacillus cirulans F-2의 Bacteriophage-like 입자)

  • 김철호;권석태;이대실;타니구치하지메
    • Microbiology and Biotechnology Letters
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    • v.18 no.3
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    • pp.221-226
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    • 1990
  • To detect prophages and bacterioeins, twenty strains of Bacillus circulans were treated with mitomycin C. The resulted lysates were subjected to electron microscopy, and also examined for killing and plaque-forming activities. Fifteen strains showed killing activity on two or more strains of Bacillue circulans. Killing agents were centrifuged in linear 5 to 20% sucrose gradient, and studied with electron microscopy which revealed the presence of particles.They looked morphologically like phage tail of 190 nm long with fiber (FA9, FA5) or without fiber (FA1, FA6), T even phage-like particle with a head of 50 nm in diameter and a tail of 140 nm long (FA7), or T7 phage-like particle with a head of 70 nm in diameter and a tail of 20 nm long (FA17). The killing agent of FA17 showed phage-forming activity on several strains different from killing sensitive strains of Bacillus circulans.

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Purification and Characterization of Phytoferritin

  • Oh, Suk-Heung;Cho, Sung-Woo;Kwon, Tae-Ho;Yang, Moon-Sik
    • BMB Reports
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    • v.29 no.6
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    • pp.540-544
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    • 1996
  • Ferritins from germinated pumpkin seeds were isolated by ammonium sulfate precipitation (0.55 saturation), ion-exchange chromatography on DEAE-cellulose, and gel filtration chromatographies on Sephacryl S-300 and Sephadex G-100. Pumpkin ferritin contains less iron than soybean ferritin. Pumpkin ferritin cross-reacted with anti-soybean ferritin antiserum made in rabbit, and showed two distinct antibody reactive bands, both of equal intensity. The pumpkin ferritins corresponding to the two bands were separable by centrifugation in a sucrose gradient (20~50%). The molecular weights of the native pumpkin ferritins based on the estimation of sucrose gradient centrifugation, gel filtration on Sephacryl S-300 and non-denaturing polyacrylamide gel electrophoresis appeared to be: 530~580 KD (the large molecular weight pumpkin ferritin) and 330-360 KD (the small molecular weight pumpkin ferritin) The large molecular weight pumpkin ferritin contains less iron. Both pumpkin ferritins cross-reacted with anti-soybean ferritin antibody with a spur formation suggesting partial antigenic recognition.

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Localization of Lipoxygenase in Germinating Soybeans (발아중인 대두콩에서의 Lipoxygenase의 국재(局在))

  • Song, Young-Sun
    • Korean Journal of Food Science and Technology
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    • v.19 no.5
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    • pp.441-445
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    • 1987
  • The subcellular distribution of lipoxygenase in germinating soybean seeds (Glycine max[L.] AmSoy) was investigated by using differential centrifugation and sucrose density gradient fractionation. Most of lipoxygenase -1 and -2/3 activities was present in the supernatant fraction after differential centrifugation of homogenates prepared from three-day-old seedlings; only 1.5% of lipoxygenase activity remained in particulate fractions. The results of a sucrose density gradient fractionation (three-day-old) showed that the lipoxygenase activity coincided with acid phosphatase at the densities of 1.19, 1.23, $1.25g/cm^3$, even though most of lipoxygenase and acid phosphatase activities appeared in supernatant fractions. There was no indication that mitochondria contained any lipoxygenase activity, and it does not appear that glyoxysomes and ER contained any lipoxygenase activity either.

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Conservation of Excavated Lacquer-wares for using artificially water-soaked Lacquer-wares (인공수침 칠기를 이용한 고대칠기 보존연구)

  • Kim, Soo-Chul
    • Journal of Conservation Science
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    • v.21
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    • pp.49-58
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    • 2007
  • Among the treatment results of test samples of the antique lacquer-ware, the treatment with PEG#3,350 40% solution displayed excellent effect with low shrinkage ratio; in weight gain the treatment with Sucrose 19%+Glycerin 1%(t-butanol 5% in water) solution showed consistent increase. However during the impregnation process of Sucrose, the weight of the testing samples decreased by dehydration because the inner part of the test samples and the treatment solution showed concentration gradient. Therefore, we concluded longer impregnation period should be necessary to prevent dehydration. Since both higher and lower molecular weight treatment chemicals could penetrate into the wood of the lacquer-ware, air drying and conditioning after impregnation treatment with high concentration chemicals would be possible, as well as vacuum freeze-drying.

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Comparison of Several Procedures for the Preparation of Synaptosomal Plasma Membrane Vesicles

  • Yun, Il;Kim, Young-Shin;Yu, Seong-Ho;Chung, In-Kyo;Baik, Seung-Wan;Cho, Goon-Jae;Chung, Yong-Za;Kim, Seok-Hwan;Kang, Jung-Sook;Kim, In-Se
    • Archives of Pharmacal Research
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    • v.13 no.4
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    • pp.325-329
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    • 1990
  • Synaptosomal plasma membrane vesicles (SPMV) were isolated from fresh bovine cerebral cortex by several well-known procedures, and the best procedures was selected by enzymatic and morphological standards. The SPMV isolated by the modified procedure of Smith and Loh showed the highest purity. The specific activities of Na, K-ATPase, acetylcholinesterase and 5'-nucleotidase were about 5, 6-fold, 2, 5-fold and 3, 3-fold, respectively, enriched in the plasma membrane fraction as compared to those of the crude homogenate. Electron microscpic examination also showed that the membranes were in vesicular form.

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Oligomeric Characterization of GroESLx Chaperonin from Symbiotic X-Bacteric in Amoeba proteus

  • Jung, Gwang-Hyun;Ahn, Tae-In
    • Animal cells and systems
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    • v.6 no.3
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    • pp.227-232
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    • 2002
  • GroESLx proteins of symbiotic X-bacteria were overproduced in Escherichia coli and their structural characteristics were assayed after simple purification. The GroESx and GroELx were heat-stable at 8$0^{\circ}C$ and 5$0^{\circ}C$, respectively. After heat-treatment, GroESx was purified by DEAE Sephadex A-50 chromatography and GroELx was purified by step- and linear sucrose density gradient ultracentrifugation. Molecular masses of GroESx and GroELx were 50-80 kDa and 800 kDa, respectively, as estimated by sucrose density gradient ultracentrifugation. In chemical cross-linking analysis, subunits of GroESx were mostly cross-linked by incubation for 3 h in 0.4% glutaralde-hyde and GroESx was found to be composed of homo-heptamer subunits. Those of GroELx were cross-linked within 10 min in 0.3% glutaraldehyde and GroELx was in two stacks of homo-heptamer subunits. On the other hand, GroESx and GroELx proteins in a solution could not be cross-linked even after incubation for 3 h in 0.5% glutaraldehyde. GroELx was stable at 4-37$^{\circ}C$. In the presence of both GroESx and ATP, GroELx$_{14}$ was stable at 37$^{\circ}C$ but not at 4$^{\circ}C$ or 24$^{\circ}C$. Thus, we confirmed the oligomeric properties of GroESx$_{7}$ and GroELx$_{14}$ and their stability to heat and in the interaction with GroESx.x.