• Title/Summary/Keyword: Purpose Binding

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Binding of the His-tagged Tail Protein J of Bacteriophage Lambda with Escherichia coli K-12 (히스티딘으로 표지된 람다 박테리오파아지 꼬리 단백질 J와 대장균 K-12와의 결합)

  • Shin, Hae Ja
    • Journal of Life Science
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    • v.28 no.1
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    • pp.78-82
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    • 2018
  • Detection of pathogenic microorganisms takes several days by conventional methods. It is necessary to assess microorganisms in a timely manner to reduce the risk of spreading infection. For this purpose, bacteriophages are chosen for use as a biosensing tool due to their host specificity, wide abundance, and safety. However, their lytic cycle limits their efficacy as biosensors. Phage proteins involved in binding to bacteria could be a robust alternative in resolving this drawback. Here, a fragment of tail protein J (residues 784 to 1,132) of phage lambda fused with 6X His-tag (6HN-J) at its N-terminus was cloned, overexpressed, purified, and characterized for its binding with microorganisms. The purified protein demonstrated a size of about 38 kDa in sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) and bound with anti-His monoclonal antibodies. It bound specifically to Escherichia coli K-12, and not Salmonella typhimurium, Bacillus subtilis, or Pseudomonas aeruginosa in dot blotting. Binding of the protein to E. coli K-12 inhibited about 50% of the in vivo adsorption of the phage lambda to host cells at a concentration of $1{\mu}g/ml$ 6HN-J protein and almost 100% at $25{\mu}g/ml$ 6HN-J. The results suggest that a fusion viral protein could be utilized as a biosensing element (e.g., protein chips) for detecting microorganisms in real time.

Association of Mannose Binding Lectin Gene Polymorphisms with the Development of Kawasaki Disease: A Pilot Study (Mannose Binding Lectin 유전자 다형성과 가와사끼병 발병의 연관성에 관한 Pilot 연구)

  • Choi, Eun Hwa;Kim, Hee Sup;Lee, Hoan Jong;Choi, Jung Yun
    • Pediatric Infection and Vaccine
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    • v.15 no.2
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    • pp.195-201
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    • 2008
  • Purpose : We hypothesized that the mannose binding lectin gene (MBL2), a key molecule of innate immunity may contribute to the development of Kawasaki disease (KD) in early childhood. This study was performed to investigate the polymorphisms of MBL2 and the risk of developing KD in Korean children. Methods : The study subjects were 112 children with KD who were admitted to the Seoul National University Bundang Hospital between October 2003 and March 2005. The control subjects consisted of 224 anonymous, healthy Korean blood donors. Extracted genomic DNA was amplified for codon 54 of MBL2 exon 1 and alleles (a and b) were assigned via sequencing analysis. The frequency of the alleles of the MBL2 exon 1 was compared between the case and control groups. Results : The median age of patients was 27 months (range, 3 months-7 years), 45.5% were <24 months of age and 54.5% were ${\geq}2$ years. The genotype distribution reached Hardy-Weinberg equilibrium in both cases and control subjects. In the cases with KD, the genotypic frequencies of codon 54 polymorphisms were 67.9% for aa, 29.5% for ab, and 2.6% for bb. There were no significant differences in the overall distribution of the polymorphisms between the cases and the control subjects. In addition, the genotype distribution was not different according to age. Conclusions : Our findings indicate that the codon 54 polymorphism of the MBL2 gene is not likely to contribute to the risk of developing KD in Korean children. Further studies on the development of coronary artery lesions with regard to MBL2 genotypes are warranted.

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A Computational Approach to Definite NPs

  • Lee, Yong-Hun
    • Korean Journal of English Language and Linguistics
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    • v.3 no.1
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    • pp.89-108
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    • 2003
  • As pronouns are resolved with their antecedents, definite NPs may enter into the anaphora-antecedent relations with indefinite NPs. This paper is to provide faster and more efficient computational algorithms by which definite NPs are resolved effectively, For this purpose, this paper extends Chierchia's Binding Theory in Categorial Grammar, and definite NPs are resolved with their antecedents by similar algorithms that are used to reflexive resolution. In these algorithms, the relations between indefinite NP and definite NP are represented with λ-expressions, and definite NPs are resolved with their antecedent by λ-conversions.

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The Evaluation of Factors which influence Binding Efficiency of Modified in Vivo Erythrocyte Labeling Technique (변형 체내 표지법에 의한 적혈구 표지시 결합효율에 영향을 미치는 인자 평가)

  • Seo Han-Kyung
    • Journal of The Korean Radiological Technologist Association
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    • v.30 no.1
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    • pp.63-69
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    • 2004
  • Purpose : We underwent this study to evaluate the factors which influence labelling efficiency when modified in vivo erythrocyte labeling technique was used. Materials and methods : Thirty healthy volunteers (M : F = 19 : 11, age : 25$\pm$2yrs)

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Control of Platelet Rolling and Adhesion

  • Moskowitz, Samuel E.
    • 제어로봇시스템학회:학술대회논문집
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    • 2002.10a
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    • pp.77.1-77
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    • 2002
  • Platelets arrest bleeding and repair damaged blood vessels. The purpose of this paper is to formulate a mathematical model for the control of platelet adhesion within the vasculature consistent with experimental findings, particularly those of Frenette, Ruggeri , Savage, Yuan, Lawrence and Springer. In addition to providing some, albeit rudimentary, insight into the behavior of platelets, a numerical simulation of this theoretical model may be useful in a systematic study of pathological cases. Glycoprotein receptor complex (GPIb/V/IX), found on the platelet surface membrane, binds to the adhesive protein and ligand von Willebrand factor (vWf), located within the sub-endothelium. The binding...

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The synergistic regulatory effect of Runx2 and MEF transcription factors on osteoblast differentiation markers

  • Lee, Jae-Mok;Libermann, Towia A.;Cho, Je-Yoel
    • Journal of Periodontal and Implant Science
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    • v.40 no.1
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    • pp.39-44
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    • 2010
  • Purpose: Bone tissues for clinical application can be improved by studies on osteoblast differentiation. Runx2 is known to be an important transcription factor for osteoblast differentiation. However, bone morphogenetic protein (BMP)-2 treatment to stimulate Runx2 is not sufficient to acquire enough bone formation in osteoblasts. Therefore, it is necessary to find other regulatory factors which can improve the transcriptional activity of Runx2. The erythroblast transformation-specific (ETS) transcription factor family is reported to be involved in various aspects of cellular proliferation and differentiation. Methods: We have noticed that the promoters of osteoblast differentiation markers such as alkaline phosphatase (Alp), osteopontin (Opn), and osteocalcin (Oc) contain Ets binding sequences which are also close to Runx2 binding elements. Luciferase assays were performed to measure the promoter activities of these osteoblast differentiation markers after the transfection of Runx2, myeloid Elf-1-like factor (MEF), and Runxs+MEF. Reverse-transcription polymerase chain reaction was also done to check the mRNA levels of Opn after Runx2 and MEF transfection into rat osteoblast (ROS) cells. Results: We have found that MEF, an Ets transcription factor, increased the transcriptional activities of Alp, Opn, and Oc. The addition of Runx2 resulted in the 2- to 6-fold increase of the activities. This means that these two transcription factors have a synergistic effect on the osteoblast differentiation markers. Furthermore, early introduction of these two Runx2 and MEF factors significantly elevated the expression of the Opn mRNA levels in ROS cells. We also showed that Runx2 and MEF proteins physically interact with each other. Conclusions: Runx2 interacts with MEF proteins and binds to the promoters of the osteoblast markers such as Opn nearby MEF to increase its transcriptional activity. Our results also imply that osteoblast differentiation and bone formation can be increased by activating MEF to elicit the synergistic effect of Runx2 and MEF.

Experimental study on the periodontal regenerative capacity of moldable synthetic peptide domain gel in degree III furcation defect of beagles (성견의 3급 이개부 병변에서 성형성 합성 펩타이드 젤의 치주재생 능력에 관한 실험적 연구)

  • Kim, Jeong-Beom;Park, Yoon-Jeong;Lee, Sang-Cheol;Kim, Tae-Il;Seol, Yang-Jo;Lee, Yong-Moo;Gu, Young;Rhyu, In-Chul;Han, Soo-Boo;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.38 no.4
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    • pp.621-628
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    • 2008
  • Purpose: Osteopontin is one of the major non-collagenous protein of hard tissue. Use of peptide domain of biologically active protein has some advantages. The objective of this experimental study is evaluation of periodontal regenerative potency of synthetic peptide gel which containing collagen binding domain of osteopontin in the degree III periodontal defect of beagle dogs. Material and Methods: Experimental degree III furcation defect was made in the mandibular third and fourth premolar of beagles. Regenerative material was applied during flap operation. 8 weeks after regenerative surgery, all animals were sacrificed and histomorphometric measurement was performed to calculate the linear percentage of the new cementum formation and the volume percentage of new bone formation. Result: The linear percent of new cementum formation was 41.6% at control group and 67.1% at test group and there was statistically significant difference. The volume percent of new bone formation was 52.1% at control group and 58.9% at test group. Conclusion: As the results of present experiment, synthetic peptide gel containing collagen binding domain of osteopontin significantly increase new bone and cementum formation in the degree III furcation defect of canine mandible.

The Effect of a Potent Oxytocin Antagonist, Antag I, on In Vitro Uterine Contractions in Response to Exogenous Oxytocin and on Uterine Oxytocin Receptor Number and Affinity (옥시토신 길항제, Antag I이 옥신토신 투여에 따른 자궁수축과 자궁의 옥시토신 수용체 수 및 친화력에 미치는 영향)

  • ;C. Warnell;G. Flouret;L. Wilson Jr.
    • Korean Journal of Animal Reproduction
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    • v.18 no.2
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    • pp.95-99
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    • 1994
  • The purpose of the present study was to determine the in vivo effect of oxytocin antagonist-I(Al) on uterine oxytocin receptor number (Rn) and/or binding affinity (Kd) in the estrous rat. Anesthetized rats were given a bolus infusion of control or 5${\mu}\textrm{g}$ of AI and sacri-ficed 0.5 and 4 hours later. The uterine tissue was removed, trimmed and frozen. Membrane oxytocin receptors were isolated after homogenization of uterine tissue and differential ultracentrifugation. The oxytocin receptor assay was performed by saturation with cold oxytocin competion with a high specific activity oxytocin antagonist. Rn and Kds were determined by nonlinear curve fitting methods. No differences(p>0.05) between the AI and control treated animals in either oxytocin receptor number or binding affinity was detected in this study. These data suggest that the major mode of action of AI is via competitive inhibition at the uterine oxytocin receptor and not by altering receptor number or binding affinity.

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The biological effects of fibrin-binding synthetic oligopeptides derived from fibronectin on osteoblast-like cells

  • Kim, Yun-Jeong;Park, Yoon-Jeong;Lee, Yong-Moo;Rhyu, In-Chul;Ku, Young
    • Journal of Periodontal and Implant Science
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    • v.42 no.4
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    • pp.113-118
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    • 2012
  • Purpose: The aim of this study was to investigate the effects of synthetic fibronectin (FN) fragments, including fibrin binding sites from amino-terminal FN fragments containing type I repeats 1 to 5, on osteoblast-like cell activity. Methods: Oligopeptides ranging from 9 to 20 amino acids, designated FF1, FF3, and FF5, were synthesized by a solid-phase peptide synthesizing system, and we investigated the effects of these peptides on cell attachment and extent of mineralization using confocal microscopy, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays, and Alizarin red S staining. Results: FF3 and FF5 peptides increased the number of attached human osteoblastic cells, and FF3 administration led to prominent cell spreading. Mineralization was increased in FF3 and FF5 compared to FF1 and the untreated control. Conclusions: Taken together, it can be concluded that the fibrin-binding oligopeptides FF3 and FF5 enhanced cell attachment and mineralization on osteoblast-like cells. These results indicate that FF3 and FF5 have the potential to increase osteoblast-like cell activity. Performing an in vivo study may provide further possibilities for surface modification of biomimetic peptides to enhance osteogenesis, thus improving the regeneration of destroyed alveolar bone.

Effect of E-box and E2F Binding Site on Transcriptional Activity in MCM Promoter (MCM promoter에서 E-box와 E2F 결합부위가 전사활성에 미치는 영향)

  • 권현주
    • Journal of Life Science
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    • v.14 no.5
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    • pp.732-740
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    • 2004
  • MCM proteins are essential for eukaryotic DNA replication, playing roles in the initiation and elongation of DNA replication. MCM proteins expression is much higher in malignant tissues than normal tissues. Several reports have indicated the usefulness of MCM proteins as markers of cancer cells in histopathological diagnosis. However, the cause of enhanced expression of MCM proteins in cancer cells remain to be clarified. The purpose of this study is to examine the relative transcriptional activities of human mcm gene promoters in cancer and normal cells. The minimal promoter region required for transcription of a luciferase reporter gene was contained an E-box and one E2F site. In addition, luciferase activities from mcm7 and mcm2 promoter/luciferase gene reporter constructs were significantly increased in cancer cells at 8 times compared with normal cells. E-box and E2F binding site in the promoter of mcm genes are responsible for different mechanism of transcription regulation on the cellular environment.