• Title/Summary/Keyword: KDA

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Purification and NMR studies on Phosphatase domain of UBLCP1

  • Oh, Hyo-Sun;Ko, Sung-Geon;Moon, Sun-Jin;Shin, Hang-Cheol;Lee, Weon-Tae
    • Journal of the Korean Magnetic Resonance Society
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    • v.13 no.2
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    • pp.126-134
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    • 2009
  • UBLCP1 is composed of Ubiquitin Like domain and RNA Polymerase II Phosphatase I domain. Phosphatase domain (25.9KDa) has been cloned into the E.coli using pET32a vector with TEV protease cleavage site and successfully purified as a monomer using affinity chromatography and histidine tag was cleaved with TEV protease for structural studies. Our results indicated that the Phosphatase domain showed well-defined folded structure based on data from one-dimensional and two-dimensional NMR spectroscopy. Data form circular dichroism also suggested that Phosphatase domain consisted of both ${\alpha}$ -helix and ${\beta}$ -sheet. This information will be used for detailed structural study of UBLCP1.

Binding of Methylene Blue to two types of water soluble polymer and its removal by polyelectrolyte enhanced ultrafiltration

  • Mansour, Nadia Cheickh;Ouni, Hedia;Hafiane, Amor
    • Membrane and Water Treatment
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    • v.9 no.2
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    • pp.87-94
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    • 2018
  • The interactions of water soluble polymers with dye are studied by ultrafiltration using a molecular weight cut off of 10 KDa regenerated cellulose ultrafiltration membrane. Two water-soluble polymers, namely Poly (Sodium-4 Styrenesulfonate) (PSS) and Poly (Vinyl Alcohol) (PVA) were selected for this study. The effects of process parameters, such as, polyelectrolyte concentrations, transmembrane pressure, ionic strength and pH of solution on dye retention and permeation flux were examined. PSS enhanced ultrafiltration achieved dye retention as high as 99% as a result of complexation between polyanion containing aromatic groups and cationic dye. This result was confirmed by the red shift. The retention of dye decreases as the salt concentration increases, a high retention was obtained at pH above 4. However, in case of PVA, relatively low retention (50%) was observed. Ionic strength and pH has no significant effect on the removal of MB. The permeate flux depended slightly on polyelectrolytes concentrations, transmembrane pressure, salt concentration and pH.

Molecular Identification and Expression of Myosin Light Chain in Shortspine Spurdog (Squalus mitsukurii)

  • Kim, Soo Cheol;Sumi, Kanij Rukshana;Sharker, Md Rajib;Kho, Kang Hee
    • Journal of Marine Life Science
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    • v.3 no.1
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    • pp.1-8
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    • 2018
  • Myosin is considered as the vital motor protein in vertebrates and invertebrates. Our present study was conducted to decipher the occurrence of myosin in dog fish (Squalus mitsukurii). We isolated one clone containing 979 bp cDNA sequence, which consisted of a complete coding sequence of 453 bp and a deduced amino acid sequence of 150 amino acids from the open reading frame with molecular weight, isoelectric point and aliphatic index are 16.72 Kda, 4.49 and 78.00, respectively. It contained 428 bp long 3' UTR with single potential polyadenylation signals (AATAAA). The predicted EF CA2+ binding domains were identified in residue 6-41, 83-118 and 133-150. A BLAST search indicates this protein exhibits a strong similarity to whale shark (Rhincodon typus) MLC3 (91% identical) and also house mouse (Mus musculus) MLC isoform 3f (81% identical). Phylogenetic analysis revealed that this protein is a MLC 3 isoform like protein. This protein also demonstrates highly conserved region with other myosin proteins. Homology modeling of S. mitsukuri was performed using crystal structure of Gallus gallus skeletal muscle myosin II based on high similarity. Reverse transcription-polymerase chain reaction (PCR), quantitative PCR results exhibits dogfish myosin protein is highly expressed in muscle tissue.

Analysis of Thermotolerance in Hot Pepper Using the Antiserum Against Carrot HSP17

  • Hwang, Eun-Young;Hwang, Cheol-Ho;Yoo, Il-Woong
    • Journal of Plant Biotechnology
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    • v.3 no.1
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    • pp.7-12
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    • 2001
  • An antiserum against the carrot HSP17 (17 KDa heat shock protein) was raised using the HSP17 purified after being expressed in a recombinant E.coli in order to develop an assay system for thermotolerance in crops. The DCHsp17.7 including the coding sequence corresponding to a carrot HSP17 protein was recombined within pET-32(b) vector and achieved a maximum expression in 4 hours after an induction in E.coli. The purified DCHsp17.7 was used as an antigen to generate the corresponding antibody. The polyclonal antiserum was confirmed for it's specificity only to the low molecular weight (1mw) HSP. Besides, the possibilities to use the antiserum to interact with 1mwHSPs from other plants such as rice, cucumber, tomato, and hot pepper were examined to be plausible. To reveal any specific correlation between the amounts of 1mwHSP expressed upon HS conditions and an acquisition of thermotolerance two different approaches have been applied. first, it has been shown that only the pre-HS conditions inducing the synthesis of HSP17 allowed for the seedlings to achieve an thermotolerance and to survive the following lethal condition. Second, a western analysis using 15 different collected lines of hot peppers was performed to distinguish each other in terms of the amount of 1mwHSP. The results indicated that all 14 hot pepper lines were able to synthesize HSPs in response to an exposure to HS conditions and the amounts of the proteins synthesized at different HS temperatures were variable among the lines. There are several different patterns of 1mwHSP synthesized as a function of temperature increase observed and their correlation to physiological aspects of thermotolerance remains to be analyzed.

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The Study of Awareness and Practice of Korean Dietitians in Food Exchange Lists , Serving Size and Dietary Guidelines (우리 나라의 식품교환표 , 식품의 서어빙 분량 , 식사지침에 대한 영양사들의 인지도 및 실천에 관한 조사 연구)

  • Lee, Yeong-Nam;No, Seong-Yun
    • Journal of the Korean Dietetic Association
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    • v.7 no.1
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    • pp.9-18
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    • 2001
  • Dietitians in various fields have used food exchange lists for food preparations. However it seems that the present food exchange lists are complicated, thus they cause many problems for dietitians to use in the fields. Therefore this study evaluated to the extent of awareness and utilizations of KDA food exchange lists in dietitians and also collected dietitian's opinions for revising food exchange lists such as serving size, serving calories, and for unifying food guidelines and dietary and dietary guidelines for Korean to one simple guideline. 192 dietitians who presently work in urban and rural areas were recruited and data based on survey were collected. As results, most of dietitians(87.5%) knew well about food exchange lists, but only 7.8% of them always would use food exchange lists for menu planning, 56.3% of dietitians did not use it at all and 34.4% occasionally use it. And 88.0% of dietitians wanted to revise food exchange lists totally or partially, 69.8% of dietitians hoped to amend various calories per one serving in food exchange lists to one serving calorie. The desirable on serving calorie was selected as 100kcal(51%) or 50kcal(38%) by dietitians. The dietitians in this study understood very well dietary guidelines(86.5%) and food guidelines(88.5%) for Korean, and 66.1% dietitians wanted to unify both guidelines. In case of unification of guidelines, dietitians answered that 7-8 items(30%) or 5-6 items(27%) should be included in guideline. In the question about reference value for daily allowance, most dietitians(56%) satisfied with the present various reference values for various generation while 28% of dietitians wanted to change to have one reference value (standard with 2000kcal, adult female). This study will provide basic informations for revising or adjustment of food exchange list and dietary or food guidelines for Korean.

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Pretreatment Effect of Running Exercise on HSP70 and DOX-Induced Cardiotoxicity

  • Shirinbayan, Vahid;Roshan, Valiollah Dabidi
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5849-5855
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    • 2012
  • Objective: The purpose of this study was to determine pretreatment effects of moderate-term endurance training before the various dosages (10 and $20{_{mg.kg}}^{-1}$) of DOX on a heat shock protein ($HSP_{70kda}$) and cardiotoxicity in heart tissue. Methods: Forty-eight male rats were randomly assigned to nontraining (NT) and training (T) groups and three subgroups; $DOX{_{10mg.kg}}^{-1}$ and $DOX{_{20mg.kg}}^{-1}$ and saline treatment. The training program included treadmill running between 25-39 min/day and 15-17 m/min, 5 days/wk for 3 wk. Result: DOX administration, in particularly with $20{_{mg.kg}}^{-1}$, caused up-regulation of oxidants and cardiac damage (MDA, CK, CPK-MB and CK/CPK-MB) and down-regulation of cardioprotection ($HSP_{70}$, SOD) markers, as compared to NT+saline group. Pretreatment effect of treadmill running endurance exercise in the presence of DOX with $10{_{mg.kg}}^{-1}$ caused a significant increase in $HSP_{70}$, SOD and a significant decrease in MDA and insignificant decrease in CK, CPK-MB and CK/CPK-MB, in comparison $T+DOX_{10}$ with $NT+DOX_{10}$ group. However, there was no significant difference between $T+DOX{_{10mg.kg}}^{-1}$ and $T+DOX{_{20mg.kg}}^{-1}$ in the aforesaid markers. Conclusion: Dox-induced cardiotoxicity is related to oxidative stress. Our study suggests that pretreatment with endurance exercise may be considered as a potentially useful strategy to improve myocardial tolerance against single dose DOX-induced oxidative damage.

Tissues and Plasma Proteins of Hemiculter eigenmanni in Muddy Water of Imha Reservoir (임하호 탁수역에 서식하는 치리 (Hemiculter eigenmanni)의 조직과 혈장단백질)

  • Lee, Chung;Shin, Myung-Ja;Lee, Jong-Eun;Seo, Eul-Won
    • Korean Journal of Environmental Biology
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    • v.24 no.3
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    • pp.213-220
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    • 2006
  • Present study aims to investigate the effect of muddy water on various tissues and plasma proteins of Hemiculter eigenmanni in Imha reservoir. The gills in muddy water were shown to have clubbing in secondary lamellae and edema in primary lamellae, respectively. The size of glomerula in kidney was smaller than that of control. Scanning electron micrographs of gills revealed muddy debris and parasites attached between primary and secondary lamellae, suggesting that muddy water possibly involves to decrease in respiratory rate. By using SDS-PAGE, plasma protein bands displayed both quantitative and qualitative changes and ferritin activities were shown more stronger and Fe-3 ferritin band appeared to be specific to plasma of muddy water.

Proteomic Changes by Acupuncture Stimulation at HT7 in the Hippocampus of Rat Pups (신문혈 자침이 어린 백서 해마의 단백질 발현에 미치는 영향)

  • Bae, Chang-Hwan;Kim, Dong-Soo;Kim, Seung-Tae
    • Korean Journal of Acupuncture
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    • v.29 no.2
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    • pp.260-270
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    • 2012
  • Objectives : Hippocampus, a region of temporal lobe, plays an important role in the pathogenic mechanisms of brain diseases such as Alzheimer's disease, depression and temporal lobe epilepsy. This research is designed to investigate hippocampal changes after acupuncture stimulation at Shinmun(HT7) using 2-dimensional gel electrophoresis(2-DE). Methods : On postnatal-day 15, rat pups were randomly devided into Normal(NOR) or HT7 group. All of Pups kept with their mothers for 7 days, but pups in HT7 group received acupuncture stimulation at HT7 daily. On postnatal-day 21, hippocampus of each rat pup was dissceted 30 minutes after last acupuncture stimulation and the protein expressions were investigated using 2-DE. Results : After acupuncture stimulation at HT7, expression of 20 proteins were significantly increased. Succinate semialdehyde dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase-like, transketolase, aconitate hydratase and phosphoglucomutase-1 were related to glucose methabolism. Eukaryotic initiation factor(eIF) 4A-II, eIF 4A-III, mitochondrial Tu translation elongation factor and chain A of crystal structure of the 70-Kda heat shock cognate protein involve in the protein synthesis in ribosome. Tubulin ${\beta}$-4 chain, tubulin T ${\beta}$-15 and tubulin ${\alpha}$-1B chain comprise cytoskeleton. Glutathione S-transferase(GST) ${\omega}$-1, GST P and GST Yb-3 can reduce oxidative stress. ${\beta}$-soluble N-ethylmaleimide-sensitive fusion protein attachment protein is required for vesicular transport between the endoplasmic reticulum and the Golgi apparatus, glycerol-3-phosphate dehydrogenase plays a major role in lipid biosynthesis, creatine kinase U-type catalyses the conversion of creatine and consumes adenosine triphosphate to create phosphocreatine and adenosine diphosphate. Platelet-activating factor acetylhydrolase IB subunit alpha and voltage depedent anion-selective channel protein 2 were also increased. Conclusions : The results suggest that acupuncture stimulation at HT7 may enhance glucose and lipid metabolism, protein synthesis, cytoskeletal substance and anti-oxidative stress in hippocampus.

Production of Glucoamylase from Hybrid Constructed by Intergenic Nuclear Transfer between Saccharomycopsis sp. and Saccharomyces sp. (핵전이법에 의해 형성된 Saccharomycopsis 속과 Saccharomyces 속의 잡종에서 glucoamylase 생산에 관한 연구)

  • 양영기;임채영;김종권;문명님;이영하
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.182-188
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    • 2001
  • The glucoamylase was purified from the induced culture filtrate of hybrid between Saccharomycopsis sp. and Saccharomycopsis sp. made by nuclear transfer and characterized for some enzyme properties. The enzymewas purified 76-fold in an overall yield of 16% from the culture medium by ammonium sulfate fractionation,Sephadex G-150 gel permeation chromatography and DEAE-Sephadex A-50 ion exchage chromatography.The molecular weight of the purified glucoamylase was estimated to be 57.5 KDa on SDS-polyacrylamidegel electrophoresis and Sephadex G-150 gel permeation chromatography. The purified enzyme was active atpH-5.0 and $40^{\circ}C$. The Km value for soluble starch was 2.6 mg/ml. The enzymatic activity was stimulated inthe presence of TEX>$Ca^{2+}$, EDTA, $Co^{2+}$, $Mg^{2+}$, and $Mn^{2+}$

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Expression Analysis of the Mx Gene and Its Genome Structure in Chickens

  • Yin, C.G.;Du, L.X.;Li, S.G.;Zhao, G.P.;Zhang, J.;Wei, C.H.;Xu, L.Y.;Liu, T.;Li, H.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.7
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    • pp.855-862
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    • 2010
  • Among the known interferon-induced antiviral mechanisms, the Mx pathway is one of the most powerful pathways. The Mx protein has direct antiviral activity and inhibits a wide range of viruses by blocking an early stage of the viral replication cycle. Cloning, characterization, and expression of Mx in vivo and in vitro have been conducted. The chicken Mx gene spans 21 kb and is made up of 14 exons and 13 introns, of which the promoter region was analyzed. The real-time PCR results showed that Mx expression was increased in chicken embryo fibroblasts (CEF) after 12- and 24-h induction with polyI: C. Induction of Mx expression by poly I: C in vivo revealed tissue-specific patterns among the chicken tissues tested. A trace expression of Mx was detected in healthy chicken liver tissues from adult chickens without inducement; the expression levels in the liver, heart, and gizzard were higher than in the muscle and kidney. This is the first report to demonstrate the expression of a glutathione-S-transferase-tagged-Mx fusion protein of 75 KDa, as well as the biological activity tested by SDS-PAGE and western blotting.