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Studies on the Isolation of Lysozyme from the Egg White and the Determination of It's Bacteriolytic Activities (난백(卵白) Lysozyme의 분리법(分離法) 및 용균활성(溶菌活性)의 측정(測定)에 관(關)한 연구(硏究))

  • Her, Yong;Oh, Hong Rock
    • Korean Journal of Agricultural Science
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    • v.14 no.2
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    • pp.272-285
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    • 1987
  • In order to obtain the data for the effective separation and purification of lysozyme from egg white, optimal conditions for the homogenization of egg white and lysozyme activities of some fresh hen eggs were examined. The recovery and purity of lysozyme isolated by the direct crystallization method, the adsorption with Bentonite and the batch method with Duolite were also investigated. The results obtained were summarized as follows; 1. On the homogenization of egg white, optimal stirring time and stirring rate for the measurement of the lysozyme activities were found to be 10 min. and 2,000 rpm respectively. 2. On the activities of lysozyme in the fresh hen eggs, the activities of W. Leghorn was greater than that of R. Island or Ogol fowl, and the activities of the thick white was a little greater than that of the thin white. 3. The residual lysozyme activities of the hen eggs stored at $4^{\circ}C$ was greater than that of the ones stored at $25^{\circ}C$ or $-20^{\circ}C$ over the storage periods. 4. Hen egg lysozyme recrystallized three times by the method of direct crystallization showed the recovery of 64.3% and the increase of 29 fold in specific activities. 5. By the adsorption method with bentonite, lysozyme in the egg white was adsorbed to 95.7%, and the elution rate of the ones adsorbed was 89.1%, and the increase in specific activities was 13 fold. 6. In the experiment exploying duolite as an adsorbent, Jysozyme in the egg white was obtained with the bound rate of 97%, the recovery rate of 84.8%, and was purified by 30fold.

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Effects of Lime and Humic Acid on the Cadmium Availability and its Uptake by Rice in Paddy Soils (논토양중 카드뮴 유효도와 수도의 흡수이행에 미치는 석회 및 Humic acid 시용효과)

  • Kim, Min-Kyeong;Kim, Won-Il;Jung, Goo-Bok;Park, Kwang-Lai;Yun, Sun-Gang;Eom, Ki-Cheol
    • Korean Journal of Environmental Agriculture
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    • v.23 no.1
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    • pp.28-33
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    • 2004
  • This study was conducted to how the effect of lime and humic acid on cadmium availability and ie uptake by plant grown in contaminated paddy soils with heavy metal. The treatment levels of lime were 2.5 and 5.0 ton/ha and that of humic acid were 1 and 2%. The contents of 0.1N HCl extractable Cd were reduced with lime and humic acid and were negatively correlated with CEC as well as soil pH. The sequential extraction procedure was used to fractionate the heavy metals in soils into the designated from exchangeable (0.5 M $KNO_3$) water soluble ($H_2O$), organically bound (0.5 M NaOH), carbonate (0.05 M $Na_{2-}$ EDTA) and sulfide/residual (4 M $HNO_3$). In soil amended with 2.5 ton/ha lime and 1% humic acia che- mical forms of Cd at tillering stage were predominant exchangeable + water soluble extractable Cd, whereas that at harvesting stage were predominant carbonate + sulfide/residual extractable Cd. The exchangeable forms of Cd in soil with lime and humic acid were negatively correlated with soil pH during the harvesting period. Total absorbed Cd of paddy rice tended to occur in the order of root > stem > leaf > brown rice. Cd contents of brown rice with lime and humic acid treatment were 0.09 and 0.08 mg/kg, respectively. That were lower than control, 0.20 mg/kg. It could be that treatment of lime and humic acid in polluted soil by heavy metals would reduce the uptake of heavy metals by piano and be a temporary method of reclamation at the highly heavy Metal contaminated soils.

Loss of Specific Sequences in a Natural Variant of Potato Proteinase Inhibitor II Gene Results in a Loss of Wound-Inducible Gene Expression (감자의 단백질 분해효소 억제제 II 유전자의 특별한 염기서열의 자연적 제거로 인한 상처 유발성 발현의 소실)

  • Thornburg, Robert W.;Park, Sang-Gyu
    • Applied Biological Chemistry
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    • v.39 no.2
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    • pp.104-111
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    • 1996
  • We have isolated several proteinase inhibitor II genes pin2 from a Russet Burbank potato DNA library. One of these, pin2T was subcloned and a 1.8 kb Xbal/Nsil insert was sequenced. This fragment contained the complete Inhibitor II gene including 965 Up of flanking DNA upstream from the gene and 200 bp of flanking DNA downstream from the gene. The open reading frame encodes a protein that is similar to other reported proteinase Inhibitor II proteins. The DNA sequence of the 5' flanking region of pin2T from -714 to +1 is highly homologous (91% identity) with that of the previously isolated wound-inducible pin2K. There are, however, four small deletions in the pin2T promoter which are located at -221 to -200, -263 to -254, -523 to -426 and -759 to -708 relative to the transcription start site of the wound-inducible pin2K. Three of these deletions map to a portion of the promoter that controls the wound-inducibility of the proteinase inhibitor genes. Chimeric genes containing the promoter of the pin2T gene linked with the both CAT and GUS were constructed and transfered into tobacco plants. Analysis of these plants indicated that pin2T is not a wound-inducible gene but is expressed at low levels. Thus, wound-inducibility is lost with the concomitant natural deletion of three small regions of the promoter. Comparision of the sequences deleted in pin2T relative to the pin2K with Genebank sequences indicates that the deleted sequences contain a motif (consensus 5'-AGTAAA-3') that is found in many other wound-inducible genes but not easily found in the published promoter sequences of other plant genes. Nuclear proteins from unwounded and wounded potato leaves were bound to the proximal promoter region, downstream of the 5'-AGTAAA-3', of pin2T. The comparison of the pin2T gone with the pin2K gene indicates that the natural internal promoter deletions are likely responsible for loss of the wound-inducible phenotype in the pin2T gene.

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Dominant-species Variation of Soil Microbes by Temperate Change (온도변화에 기인한 토양미생물 우점종의 변화에 관한 연구)

  • Park, Kap-Joo;Lee, Byeong-Chol;Lee, Jae-Seok;Park, Chan-Sun;Cho, Myung-Hwan
    • Korean Journal of Environmental Biology
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    • v.29 no.1
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    • pp.52-60
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    • 2011
  • Today, the weather is changing continually, due to the progress of global warming. As the weather changes, the habitats of different organisms will change as well. It cannot be predicted whether or not the weather will change with each passing day. In particular, the biological distribution of the areas climate change affects constitutes a major factor in determining the natural state of indigenous plants; additionally, plants are constantly exposed to rhizospheric microorganisms, which are bound to be sensitive to these changes. Interest has grown in the relationship between plants and rhizopheric microorganisms. As a result of this interest we elected to research and experiment further. We researched the dominant changes that occur between plants and rhizospheric organisms due to global warming. First, we used temperature as a variable. We employed four different temperatures and four different sites: room temperature ($27^{\circ}C$), $+2^{\circ}C$, $+4^{\circ}C$, and $+6^{\circ}C$. The four different sites we used were populated by the following species: Pinus deniflora, Pinus koraiensis, Quercus acutissima, and Alnus japonica. We counted colonies of these plants and divided them. Then, using 16S rRNA analysis we identified the microorganisms. In conclusion, we identified the following genera, which were as follows: 10 species of Bacillus, 2 Enterobacter species, 4 Pseudomonas species, 1 Arthrobacter species, 1 Chryseobacterium species, and 1 Rhodococcus species. Among these genera, the dominant species in Pinus deniflora was discovered in the same genus, but a different species dominated at $33^{\circ}C$. Additionally, that of Pinus koraiensis changed in both genus and species which changed into the Chryseobacrterium genus from the Bacilus genus at $33^{\circ}C$.

Characterization on the Behavior of Heavy Metals and Arsenic in the Weathered Tailings of Songcheon Mine (송천광산의 풍화광미 내 중금속 및 비소 거동 특성)

  • Lee, Woo-Chun;Kim, Young-Ho;Cho, Hyen-Goo;Kim, Soon-Oh
    • Journal of the Mineralogical Society of Korea
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    • v.23 no.2
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    • pp.125-139
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    • 2010
  • Behavior of heavy metals and arsenic in the tailings of Songcheon Au-Ag mine was characterized via both mineralogical and geochemical methods. Mineral composition of the tailings was investigated by X-ray diffractometry, energy-dispersive spectroscopy, and electron probe micro-analyzer (EPMA) and total concentrations of heavy metals and arsenic and their chemical forms were analyzed by total digestion of aqua regia and sequential extraction method, respectively. The results of mineralogical study indicate that the tailings included mineral particles of resinous shape mainly consisting of galena, sphalerite, pyrite, quartz, and scorodite, and specifically socordite was identified in the form of matrix. EPMA quantitative analyses were performed to evaluate the weatherability of each mineral, and the results suggest that it decreased in the sequence of arsenopyrite > galena > sphalerite > pyrite. The weathering pattern of galena was observed to show distinctive zonal structure consisting of secondary minerals such as anglesite and beudantite. In addition, almost all of arsenopyrite has been altered to scorodite existing asmatrix and galena, sphalerite, and pyrite which have lower weatherability than arsenopyrite were identified within the matrix of scorodite. During the process of alteration of arsenopyrite into scorodite, it is likely that a portion of arsenic was lixiviated and caused a great deal of detrimental effects to surrounding environment. The results of EPMA quantitative analyses verify that the stability of scorodite was relatively high and this stable scorodite has restrained the weathering of other primary minerals within tailings as a result of its coating of mineral surfaces. For this reason, Songcheon tailings show the characteristics of the first weathering stage, although they have been exposed to the surface environment for a long time. Based on the overall results of mineralogical and geochemical studies undertaken in this research, if the tailings are kept to be exposed to the surface environment and the weathering process is continuous, not only hazardous heavy metals, such as lead and arsenic seem to be significantly leached out because their larger portions are being partitioned in weakly-bound (highly-mobile) fractions, but the potential of arsenic leaching is likely to be high as the stability of scorodite is gradually decreased. Consequently, it is speculated that the environmental hazard of Songcheon mine is significantly high.

Brain-expressed X-linked 2 Binds to Kinesin Superfamily Protein 3A (Brain-expressed X-linked (Bex) 2와 heterotrimeric kinesin-2의 KIF3A와의 결합)

  • Kim, Mooseong;Jeong, Young Joo;Park, Sung Woo;Seo, Mi Kyoung;Kim, Sang Jin;Lee, Won Hee;Urm, Sang-Hwa;Lee, Jung Goo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.32 no.3
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    • pp.189-195
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    • 2022
  • Kinesin-2 comprises two subfamilies of the heterotrimeric or homodimeric motors found in mammalian cells. Heterotrimeric kinesin-2 consists of kinesin superfamily proteins (KIFs) 3A and 3B and kinesin-associated protein 3 (KAP3), which is a molecular motor protein that moves along microtubules. It plays diverse roles in cargo transport, including anterograde trafficking in cilia, and interacts with many different cargoes and proteins, but their binding proteins have not yet been fully identified. In this study, the yeast two-hybrid assay was used to identify the proteins that interact with the cargo-binding domain (CBD) of KIF3A, and an interaction between KIF3A and brain expressed X-linked 2 (Bex2) was found. Bex2 bound to the CBD-containing C-terminal tail region of KIF3A but did not interact with the same region of KIF3B or KIF5A (a motor protein of kinesin-1). KIF3A interacted with another isoform, Bex1, but did not interact with Bex3. In addition, glutathione S-transferase (GST) pull-downs showed that KIF3A specifically interacts with GST-Bex1 and GST-Bex2 but not with GST alone. When co-expressed in HEK-293T cells, Bex2 co-localized with KIF3A and co-immunoprecipitated with KIF3A and KIF3B but not KIF5B. In combination, these results suggest that Bex2 is capable of binding to heterotrimeric kinesin-2 and may serve as an adaptor protein that links heterotrimeric kinesin-2 with cargo.

Empty Time, Empty Space, and Non(非)-Place in World of Warcraft: The Accumulated Experience and the Recovery in Reality (<월드 오브 워크레프트>(World of Warcraft)의 빈 시간, 빈 공간, 비(非) 장소: 축적된 체험과 실감의 회복)

  • Song, Kyong-Won
    • Cartoon and Animation Studies
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    • s.19
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    • pp.127-143
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    • 2010
  • The purpose of this study is to aesthetically approach a game, which was popular at home, through microscopic analysis on World of Warcraft. World of Warcraft(hereunder called WOW), which improved a game method of MMORPG that had been popular conventionally at home, from several angles, is suggesting new paradigm of MMORPG game. This study aims to propose new vision on game analysis by approaching this new method from the perspective of player. For this, it divided WOW into shift method, instruction implementation method, and communication configuration method, and analyzed this, respectively, by introducing a concept of empty time, empty space, and non-place, which was borrowed from Zygmunt Bauman's "Liquid Modernity". WOW's shifting method rather extends empty time, that is, the waiting time, contrary to the conventional MMORPG, which used a method of removing. Instead of maximally reducing boring, which becomes disturbance of a game, it is what overwhelmingly enlarges time that needs to be waited, thereby being what induces a sense of time, which a player experiences daily life, to the inside of a game. WOW's instruction implementation method offers one of hugely single map instead of zone-system necessary for map loading in the discontinued form. This minutely implements even a greatly insignificant place in playing a game, thereby stirring up a sense of travel, which explores there in reality. Finally, WOW's community configuration method makes a hunting-targeted group clear, which added a concept of hunting called the raid to the existing gild system. The raid, which is a large party of being bound to the inside of Instant Dungeon, clearly gives a performance role to each of party members, thereby allowing the identity in character to be connected directly to the identity of player. Through this, the player filled the suggested 'space' with experience, thereby being able to change it into 'place' that is significant to an individual.

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Fate of the herbicide bensulfuron-methyl in a soil/rice plant microecosystem (벼 재배 microecosystem 내에서 제초제 bensulfuron-methyl의 행적)

  • Lee, Jae-Koo;Fuhr, F.;Kwon, Jeong-Wook;Ahn, Ki-Chang;Park, Ju-Hyoung;Lee, Yong-Pil
    • The Korean Journal of Pesticide Science
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    • v.8 no.4
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    • pp.299-308
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    • 2004
  • In order to elucidate the behavior of bensulfuron-methyl, a sulfonylurea herbicide, in a soil/plant microecosystem, rice plants (Oryza sativa L.) were grown for 12 weeks in the specially made stainless steel pots (17cm I.D. $\times$ 10cm H.) containing two different paddy soils treated with fresh and 13-week-aged residues of [phenyl-$^{14}C$]bensulfuron-methyl, respectively. During the aging period, the mineralization to $^{14}CO_2$ from soil A (OM, 3.59%; CEC, 7.65 $cmol^+\;kg^{-1}$; texture, sandy clay loam) and B (OM, 1.62%; CEC, 4.51 $cmol^+\;kg^{-1}$; texture, sandy loam) amounted to 6.79 and 10.15% of the originally applied $[^{14}C]$bensulfuron-methyl, respectively. The amounts of $^{14}CO_2$ evolved from the soils with fresh residues were higher than those from the soils with aged residues. At harvest after 12-week growing, $^{14}C$-radioactivity absorbed and translocated into rice plants from soils A and B containing fresh residues of bensulfuron-methyl was 1.53 and 4.40%, while 4.04 and 6.37% in the two soils containing aged residues, respectively. Irrespective of aging and soil type, the $^{14}C$-radioactivity remaining in soil ranged from 80.41 to 98.87% of the originally applied $[^{14}C]$bensulfuron-methyl. The solvent extractability of tile soils was $39.25\sim70.39%$, showing the big differences among the treatments. Most of the nonextractable soil-bound residues of $[^{14}C]$bensulfuron-methyl were incorporated into the fulvic acid fraction$(61.32\sim76.45%)$. Comparing the microbial activity of the soils with rice plants grown with that of the soils without them, the former was $1.6\sim3.0$ times higher than the latter. However, it did not correlate with the $^{14}CO_2$ evolution.

Muskelin Interacts with Multi-PDZ Domain Protein 1 (MUPP1) through the PDZ Domain (Muskelin과 multi-PDZ domain protein 1 (MUPP1) 단백질의 PDZ 도메인을 통한 결합)

  • Jang, Won Hee;Jeong, Young Joo;Choi, Sun Hee;Lee, Won Hee;Kim, Mooseong;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.25 no.5
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    • pp.594-600
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    • 2015
  • Protein-protein interactions have a critical role in the regulation of many cellular functions. Postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domain is one of domains that mediate protein-protein interactions. PDZ domains typically bind to the specific motif at the carboxyl (C)-terminal end of partner proteins. Multi-PDZ domain protein 1 (MUPP1), which has 13 PDZ domains, serves a scaffolding function for structure proteins and signaling proteins, but the cellular function of MUPP1 has not been fully elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and muskelin. Muskelin was recently identified as a GABAA receptor (GABAAR) α1 subunit binding protein and known to have a role in receptor endocytosis and degradation. Muskelin bound to the 3rd PDZ domain, but not to other PDZ domains of MUPP1. The C-terminal end of muskelin was essential for the interaction with MUPP1 in the yeast two-hybrid assay. When co-expressed in HEK-293T cells, muskelin but not the C-terminal deleted muskelin was co-immunoprecipitated with MUPP1. In addition, MUPP1 co-localized with muskelin at the same subcellular region in cells. These findings collectively suggest that MUPP1 or its interacting proteins could modulate GABAAR trafficking and turnover through the interaction with muskelin.

Development and Preliminary Evaluation of a Leukocyte Removal Aptamer Filter (압타머를 이용한 백혈구제거필터의 개발 및 예비평가)

  • Lee, Yangwon;Jung, Eun-Suk;Choi, Kyoung Young;Kim, Myung Han;Kwon, So Yong;Cho, Nam Sun;Kim, Jin Sook;Park, Han Jeong;Han, Byoung Don;Yoon, Soo-Young
    • The Korean Journal of Blood Transfusion
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    • v.23 no.2
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    • pp.107-114
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    • 2012
  • Background: Leukocyte reduction filters are widely used to prevent transfusion reactions caused by leukocytes in blood components. Commercial filters are not sufficient for removal of leukocytes for prevention of transfusion associated graft-versus-host disease; therefore, irradiation of blood components was performed using expensive equipment. Techniques using an aptamer substituted for antibody have been developed and are available in clinical areas. The purpose of this study is to develop the aptamer filter system and to evaluate its efficiency and the possibility of its clinical application. Methods: Aptamers targeted to CD45 were selected by the Postech Aptamer Initiative. The aptamer filter in which aptamers attached to beads were bound to leukocytes and removed by magnetic field was developed. Filtration of 14 units of leukoreduction-red blood provided by Korean Red Cross Blood Services was performed using aptamer filters. Leukocyte removal rate and red cell recovery rate were evaluated and bacterial culture was performed. Results: After filtration using the aptamer filters, 45.6% of leukocytes were additionally removed and the red cell recovery rate was 92.8%. No growth in the bacterial culture was observed. Conclusion: In order to apply the cell depletion technique utilizing an aptamer to blood filter system, we developed and evaluated the aptamer filter system. Through improvement of the binding efficiency of the aptamer and the filtering process, and application of the various aptamers for other different cells, we suggest that this technique can be applied in the clinical area, such as a substitution for the irradiation process for TAGVHD prevention.