• Title/Summary/Keyword: molecular weights

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Expression and Characterization of Human T-Cell Leukemia Virus Type-I Env and Gag Proteins

  • Son, Kyung-Hwa;Kim, Byong-Moon;Lee, Taik-You;Kim, Seong-Ryong;Kim, Kun-Soo;Lee, Jeong-Kug;Yang, Jai-Myung
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.311-317
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    • 1999
  • Human T-cell leukemia virus Type-I (HTLV-I) is etiologically associated with rare adult T-cell leukemia, a malignant T-cell disorder. cDNAs encoding p24 (gag), gp21(env), and pXII of HTLV-I were amplified by polymerase chain reaction (PCR) using the genomic DNA extracted from HUT102 cell line as a template. The amplified cDNAs were cloned into the Escherichia coli expression vectors and over-expression of the recombinant proteins were achieved by adding IPTG into the culture media in order to induce the promoter. The molecular weights of the recombinant p24, gp21, and pXII, determined by SDS-PAGE, were found to be approximately 28 kDa, 23 kDa, and 15 kDa, respectively. Reactivity of the recombinant proteins with human sera was tested by the immunoblot assay. The gp21 and p24 reacted against the sera obtained from HTLV-I-infected individuals but not against the sera obtained from normal persons. These results suggest that the recombinant proteins expressed in E. coli were recognized by antibodies in sera from HTLV-I infected patients. These recombinant proteins would be applicable for detecting the presence of antibodies against HTLV-I in human blood samples.

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Consistent and Specific Suppression of Mucin Release from Cultured Hamster Tracheal Surface Epithelial Cells by Poly-L-Lysine

  • Lee, Choong-Jae;Lee, Jae-Heun;Seok, Jeong-Ho;Hur, Gang-Min;Park, Ji-Sun;Bae, So-Hyun;Jang, Hyeon-Seok;Park, Sang-Cheol
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.3
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    • pp.143-147
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    • 2003
  • Poly-L-lysine (PLL) was reported to suppress mucin release from airway goblet cells during 30 min treatment period. In this study, we investigated whether PLL consistently suppresses mucin release from cultured airway goblet cells during 24 h after 30 min treatment and also specifically suppresses the release of mucin without any effects on the other releasable glycoproteins. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with $^3H$-glucosamine for 24 h and chased for 30 min in the presence of varying concentrations of PLL to assess the effects on $^3H$-mucin release and on the total elution profile of the treated culture medium. The total mucin content during 24 h after 30 min treatment of PLL was assesed to investigate the consistency of effects. PLL did not affect the release of the other releasable glycoproteins whose molecular weights were less than mucin, and decreased the total mucin content during 24 h after 30 min treatment. We conclude that PLL can specifically suppress mucin release from cultured airway goblet cells and the suppression on mucin release is consistent. This finding suggests that PLL might be used as a specific airway mucin-regulating agent by directly acting on airway mucin-secreting cells.

Isolation of a Poly (L-lactide) Degrading Bacterium and Improvement of its Degradation Capacity (Poly(L-lactide)분해 세균의 분리 및 활성 증진)

  • Kim, Mal-Nam;Park, Sang-Tae
    • Korean Journal of Environmental Biology
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    • v.25 no.3
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    • pp.260-266
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    • 2007
  • A thermophilic bacterium capable of poly (L-lactide)(PLLA) degradation was isolated from cultivating soil in Korea. The isolate was Gram positive rod-shaped bacterium, and was identified as Geobacillus caldoxylosilyticus based on the 16S rDNA sequence analysis. The strain proved to be a new PLLA degrading bacterium which has not been reported in the open literatures yet. The degradation activity of the strain was assessed in a sterilized compost inoculated with the strain under controlled compost condition at $58^{\circ}C$ for 40 days. The strain mineralized 66%, 57%, 41% and 40% of PLLA5000, PLLA11000, PLLA34000 and PLLA256000 whose weight average molecular weights were 5000, 11000, 34000 and 256000, respectively. Incorporation of 0.1% each of gelatin, yeast extract and ammonium sulfate in the compost containing PLLA256000 as a nutritional supplement raised the biodegradation activity by 27%, 13% and 10%, respectively. Increase of the inoculum size from $10^9cfu\;g^{-1}\;to\;10^{10}cfu\;g^{-1}\;and\;10^{11}cfu\;g^{-1}$ also enhanced the biodegradation activity by 14% and 20%, respectively.

Enzymatic Characteristics of Two Forms of the Purified Glucoamylase from Rhizopus oryzae (Rhizopus oryzae로부터 정제한 두가지형의 Glucoamylase의 효소적(酵素的) 특성(特性))

  • Hou, Won-Nyong;Chung, Man-Jae
    • Korean Journal of Food Science and Technology
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    • v.16 no.4
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    • pp.392-397
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    • 1984
  • These experiments were conducted to investigate general enzymatic characteristics of two forms(glucoamylase I and glucoamylase II) of the purified glucoamylase produced by Rhizopus oryzae. Molecular weights of glucoamylase I and glucoamylase II estimated by Sephadex G-100gel filtration, were approximately 101,000 and 115,000, respectively, and those estimated by SDS-polyacrylamide gel electrophoresis being 120,000 and 127,000, respectively. Isoelectric points of the above enzyme were pH 7.25 and pH 7.75. The optimum temperature was $50^{\circ}C$ and the enzyme was stable below $45^{\circ}C$. Optimum pH of both glucoamylase I and glucoamylase II was about pH 5.0. The stable pH range of them were pH 3.5-8.0 and 4.5-8.0, respectively. Michaelis constants of glucoamylase I and glucoamylase II toward souluble starch were 4.545 mg/ml and 5.560 mg/ml, respectively. $Hg^{++}$, $Pb^{++}$, p-CMB and IAA were inhibitors of glucoamylase I and $Hg^{++}$, $Mn^{++}$, p-CMB and IAA were inhibitors of glucoamylase II.

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A Study on the production of Monoclonal Antibodies against Rhodotrodula rubra (효모양진균의 단크론 생성)

  • Ryeom K.;Kim S. C.;Lee J. H.
    • Journal of environmental and Sanitary engineering
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    • v.4 no.1 s.6
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    • pp.17-28
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    • 1989
  • Having studied the production of monoclonal antibodies for developing a diagnosis medicine which shall be detected by a high-sensitivity test by using Rhodotorula rubra as a fungi-host which had been extracted through biochemical tests and follow-up examinations on Yeast-like fungi obtained from pulmonary tissues of pulmonary tuberculosis patients who had been in Kong ju National Tuberculosis Hospital from Jun. to Dec. in 1987, I. have gained such results as follows: 1. The fusion rate was influenced by feeder cell layers, cell density and time required to the cell fusion with cells in myelona subculture. 2. The fusion rate did not show any significant difference when the cell was applyed with two molecular weights, i.e., 1500 and 4000, of polyethylene glycol. 3. Fused cells after the addition of HAT selection media were bright and round, whereas unfused myelona cells and spleen cells were shrunk and granulated. 4. The cell fusion rate turned out to be about $57.2\%$(150 wells / 264 wells). 5. $10\%$(15 wells / 150 wells) of the positive reaction was detected in monoclonal antibody screening. 6. The titer which had reacted positively to Rhodotorula rubra fungal-host was 800 times in density after the gradual dilution of the produced monoclonal antibodies with Indirect ELISA method. 7. The Strongest specific reaction came out after the peroxidase labelled anti-human Immunogobulin had been applyed to Rhodotorula rubra for activating its nature after making drift with Carbonate-bicarbonate buffer (pH 9.6) and drying completely.

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Molecular Diffusion of Water in Paper (IV) - Mathematical model and fiber-phase moisture diffusivities for unsteady-state moisture diffusion through paper substrates - (종이내 수분확산 (제4보) - 종이의 비정상상태 수분확산 모델과 섬유상 수분확산 계수 -)

  • 윤성훈;박종문;이병철
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.34 no.3
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    • pp.17-24
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    • 2002
  • An unsteady-state moisture diffusion through cellulosic fibers in paper was characterized from the moisture sorption experiment and the mathematical modeling. The sorption experiment was conducted by exposing thin dry paper specimens to a constant temperature-humidity environment. Oven dried blotting papers and filter papers were used as test samples and the gains of their weights were constantly monitored and recorded as a function of sorption time. For a mathematical approach, the moisture transport was assumed to be an one-dimensional diffusion in thickness direction through the geometrically symmetric structure of paper. The model was asymptotically simplified with a short-term approximation. It gave us a new insight into the moisture uptake phenomena as a function of square root of sorption time. The fiber-phase moisture diffusivities(FPMD) of paper samples were then determined by correlating the experimental data with the unsteady-state diffusion model obtained. Their values were found to be on the order of magnitude of $10^{-6}-10^{-7}cm^2$/min., which were equivalent to the hypothetical effective diffusion coefficients at the limit of zero porosity. The moisture sorption curve predicted from the model fairly agreed with that obtained from the experiment at some limited initial stages of the moisture uptake process. The FPMD value of paper significantly varied depending upon the current moisture content of paper. The mean FPMD was about 0.7-0.8 times as large as the short-term approximated FPMD.

Membrane Permeation Characteristics and Fouling Control through the Coating of Poly(vinyl alcohol) on PVDF Membrane Surface (PVDF막 표면에 폴리비닐알코올 코팅을 통한 분리막의 투과특성 및 막오염 제어)

  • Jang, Hanna;Kim, In-Chul;Lee, Yongtaek
    • Membrane Journal
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    • v.24 no.4
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    • pp.276-284
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    • 2014
  • In this study, a hydrophobic polyvinylidene fluoride (PVDF) membrane was modified by coating neutral hydrophilic poly(vinyl alcohol). The flux of pure water was measured and then fouling test was conducted with bovin serum albumin (BSA) as model protein foulant. As a result, the experiments showed that pure water flux was decreased but anti-fouling property was significantly enhanced. Pure water flux with increasing molecular weights of the polymer was decreased and fouling resistance was enhanced. Also, Pure water flux with increasing solution concentration was decreased and fouling resistance was enhanced. It is probably due to the increase in hydrophilicity and decrease in roughness of the membrane surface, as revealed by contact angle and AFM analysis.

Protein Expression Analysis of Halobacillus dabanensis $D-8^T$ Subjected to Salt Shock

  • Feng De Qin;Zhang Bo;Lu Wei Dong;Yang Su Sheng
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.369-374
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    • 2006
  • To investigate the mechanism of salt tolerance of gram-positive moderately halophilic bacteria, two-dimensional gel electrophoresis (2-D PAGE) was employed to achieve high resolution maps of proteins of Halobacillus dabanensis $D-8^T$. Approximately 700 spots of proteins were identified from these 2-D PAGE maps. The majority of these proteins had molecular weights between 17.5 and 66 kDa, and most of them were distributed between the isoelectric points (pI) 4.0 and 5.9. Some protein spots were distributed in the more acidic region of the 2-D gel (pI <4.0). This pattern indicated that a number of proteins in the strain $D-8^T$ are acidic. To understand the adaptation mechanisms of moderately halophilic bacteria in response to sudden environmental changes, differential protein profiles of this strain were investigated by 2-D PAGE and $Imagemaster^{TM}$ 2D Platinum software after the cells were subjected to salt shock of 1 to 25% salinity for 5 and 50 min. Analysis showed 59 proteins with an altered level of expression as the result of the exposure to salt shock. Eighteen proteins had increased expression, S proteins were induced, and the expression of 33 proteins was down-regulated. Eight of the up-regulated proteins were identified using MALDI-TOF/MS and MASCOT, and were similar to proteins involved in signal transduction, proteins participating in energy metabolism pathways and proteins involved in stress.

Purification and Some Properties of Soluble Cytochrome c from Methylobacillus sp. Strain SK1 (Methylobacillus sp. Strain SK1에 존재하는 Soluble cytochrome c의 정제 및 특성)

  • 김시욱;노영태;김영민
    • Korean Journal of Microbiology
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    • v.29 no.6
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    • pp.380-386
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    • 1991
  • Three types of soluble cytochrome c were purified to homogeneity from Methylobacillus sp. strain SK1 which grows only on methanol. Cytochrome c-I was purified 58.5-fold in seven steps. Cytochrome c-II and c-III were purified 57.3- and 122.1-fold in eight steps, respectively. The molecular weights of the cytochrome c-I was determined to be 12,500, while those of the cytochrome c-II and c-III were 16,000. The isoelectric points of the c-I, c-II and c-III were found to be 8.8, 6.6, and 6.6 respectively. The spectrum of reduced cytochrome c-I showed .alpha.-, .betha.-, .gamma.-peaks at 551.4, 522.2, and 416.6nm. The peaks for c-II were found at 551.0, 521.6, and 416.5nm, while those for c-III were shown at 551.2, 521.8, and 416.0 nm. The spectra of oxidized cytochrome c-I, c-II, and c-III showed .gamma.-peak at 411.8, 409.0, and 410.2 nm, respectively. The absorption coefficients of .alpha.- and .gamma.-peak for c-I in the reduced state were determined as 47 and 197 $mM^{-1}$ $cm^{-1}$ , respectively. The coefficients of .alpha.- and .gamma.-peak for c-II were determined to be 43 and 137 $mM^{-1}$ $cm^{-1}$ , while those for c-III were 41 and 172 $mM^{-1}$ $cm^{-1}$ , respectively. The c-I and c-III were found to bind carbon monoxide.

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Preparation of Biodegradable PLGA Microspheres for Sustained Local Anesthesia and Their in vitro Release Behavior (지속적인 국소마취를 위한 생분해성 PLGA 미립구의 제조와 생체외 방출 거동)

  • 조진철;강길선;최학수;이종문;이해방
    • Polymer(Korea)
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    • v.24 no.5
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    • pp.728-735
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    • 2000
  • Fentanyl-loaded biodegradable poly(L-lactide-co-glycolide) (75 : 25 by mole ratio of lactide to glycolide, PLGA) microspheres (MSs) were prepared to study the possibility for long-acting local anesthesia. We developed the fentanyl base (FB, slightly water-soluble)-loaded PLGA MSs by means of conventional O/W solvent evaporation method. The size of MSs was in the range of 10~150 ${\mu}{\textrm}{m}$. The morphology of MSs was characterized by SEM, and the in vitro release amounts of FB were analyzed by HPLC. The lowest porous cross-sectional morphology and the highest encapsulation efficiency were obtained by using gelatin as an emulsifier. The influences of several preparation parameters, such as emulsifier types, molecular weights and concentrations of PLGA, and initial drug loading amount, etc., have been observed in the release patterns of FB. The release of FB in vitro was more prolonged over 25 days, with close to zero-order pattern by controlling the preparation parameters. We also investigated the physicochemical properties of FB-loaded PLGA MSs by X-ray diffraction and differential scanning calorimeter.

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