• Title/Summary/Keyword: Self-Affinity

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Comparison of Enzymatic Activity and Cleavage Characteristics of Trypsin Immobilized by Covalent Conjugation and Affinity Interaction (공유결합과 친화력결합에 의한 고정화 Trypsin의 효소역가와 절단특성 비교)

  • Jang, Dae-Ho;Seong, Gi-Hun;Lee, Eun-Kyu
    • KSBB Journal
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    • v.21 no.4
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    • pp.279-285
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    • 2006
  • We investigated the effects of immobilization chemistry on the yield of immobilization and the bioactivity of the immobilized enzymes. Trypsin as a model protein and macroporous polymer beads(Toyopearl AF 650M, Tosho Co., Japan) was used as a model matrix. Four methods were used to immobilize trypsin; covalent conjugation by reductive amination(at pH 10.0 and pH 4.0) and affinity interaction via streptavidin-biotin, and double-affinity interaction via biotin-streptavidin-biotin system. The covalent conjugation immobilized $3{\sim}4$ mg/ml-gel, ca. 3-fold higher than the affinity method. However, the specific activity of the covalently(pH 10.0) and affinity-immobilized trypsin(via streptavidin-biotin) are ca. 37% and 50%, respectively, of that of the soluble enzyme(on the low-molecular-weight BAPNA substrate). When the molecular size of a substrate increased, the affinity-immobilized trypsin showed higher clavage activity on insulin and BSA. This result seemed to indicate the streptavidin-biotin system allowed more steric flexibility of the immobilized trypsin in its interaction with a substrate molecule. To confirm this, we studied the molecular flexibility of immobilized trypsin using quartz crystal microbalance-dissipation. Self-assembled monolayers were formed on the Q-sensor surface by aminoalkanethiols, and gultaraldehyde was attached to the SAMs. Trypsin was immobilized in two ways: reductive amination(at pH 10.0) and the streptavidin-biotin system. The dissipation shift of the affinity-immobilized trypsin was $0.8{\times}10^{-6}$, whereas that of the covalently attached enzyme was almost zero. This result confirmed that the streptavidin-biotin system allowed higher molecular flexibility. These results suggested that the bioactivity of the immobilized enzyme be strongly dependent on its molecular flexibility.

The Relationships of Elementary School Students' Emotional Affinity toward and Intellectual Curiosity about Life, and Experiences about Nature and Life with Their Pro-Environmental Behavior (초등학생의 생물에 대한 정서적 친밀감 및 지적 호기심, 자연 및 생물에 대한 경험과 환경 친화적 행동의 관계)

  • Kim, Heung-Tae;Jeon, Min-Jeong;Kim, Jae Geun
    • Journal of Korean Elementary Science Education
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    • v.31 no.2
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    • pp.197-207
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    • 2012
  • The purpose of this study was to assess pro-environmental behavior of elementary school students by school year and gender and to investigate the relationships of their emotional affinity toward and intellectual curiosity about life, and experiences about nature and life with the pro-environmental behavior. A total of 436 elementary students participated in the study: 189 3rd grade; 247 6th grade students, and 233 boys; 203 girls. The variables were measured by using a self-report questionnaire with 5-point Likert scale items. The results showed the levels of pro-environmental behavior of the 3rd grade students and the girls were higher than that of the 6th grade and the boys, respectively. Their pro-environmental behavior was significantly related with all measured variables. However, it was found that the direct predictors of their pro-environmental behavior were the intellectual curiosity and the indirect experience, while the emotional affinity and the direct experience did not have a direct impact on the behavior. The predictive power of the intellectual curiosity and the indirect experience for their pro-environmental behavior was higher in 3rd grade than 6th grade. While the indirect experience was a stronger predictor of the boys' pro-environmental behavior than the intellectual curiosity, the curiosity was a stronger predictor of the girls.

Activation of the Mating Pheromone Response Pathway of Lentinula edodes by Synthetic Pheromones

  • Ha, Byeongsuk;Kim, Sinil;Kim, Minseek;Ro, Hyeon-Su
    • Mycobiology
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    • v.46 no.4
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    • pp.407-415
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    • 2018
  • Pheromone (PHB)-receptor (RCB) interaction in the mating pheromone response pathway of Lentinula edodes was investigated using synthetic PHBs. Functionality of the C-terminally carboxymethylated synthetic PHBs was demonstrated by concentration-dependent induction of a mating-related gene (znf2) expression and by pseudoclamp formation in a monokaryotic strain S1-11 of L. edodes. Treatment with synthetic PHBs activated the expression of homeodomain genes (HDs) residing in the A mating type locus, and of A-regulated genes, including znf2, clp1, and priA, as well as genes in the B mating type locus, including pheromone (phb) and receptor (rcb) genes. The synthetic PHBs failed to discriminate self from non-self RCBs. PHBs of the B4 mating type (B4 PHBs) were able to activate the mating pheromone response pathway in both monokaryotic S1-11 and S1-13 strains, whose B mating types were B4 (self) and B12 (non-self), respectively. The same was true for B12 PHBs in the B4 (non-self) and B12 (self) mating types. The synthetic PHBs also promoted the mating of two monokaryotic strains carrying B4-common incompatible mating types ($A5B4{\times}A1B4$). However, the dikaryon generated by this process exhibited abnormally high content of hyphal branching and frequent clamp connections and, more importantly, was found to be genetically unstable due to overexpression of mating-related genes such as clp1. Although synthetic PHBs were unable to discriminate self from non-self RCBs, they showed a higher affinity for non-self RCBs, through which the mating pheromone response pathway in non-self cells may be preferentially activated.

Development of Surface Plasmon Resonance Immunosensor through Metal Ion Affinity and Mixed Self-Assembled Monolayer

  • Lee, Si-Ra;Sim, Sang-Jun;Park, Chul-Hwan;Gu, Man-Bock;Hwang, Un-Yeon;Yi, Jong-Heop;Oh, Byung-Keun;Lee, Jin-Won
    • Journal of Microbiology and Biotechnology
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    • v.18 no.10
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    • pp.1695-1700
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    • 2008
  • An immunosensor based on surface plasmon resonance (SPR) with enhanced performance was developed through a mixed self-assembled monolayer. A mixture of 16-mercaptohexadecanic acid (16-MHA) and 1-undecanethiol with various molar ratios was self-assembled on gold (Au) surface and the carboxylic acid groups of 16-MHA were then coordinated to Zn ions by exposing the substrate to an ethanolic solution of $Zn(NO_3)_2\cdot 6H_2O$. The antibody was immobilized on the SPR surface by exposing the functionalized substrate to the desired solution of antibody in phosphate-buffered saline (PBS) molecules. The film formation in series was confirmed by SPR and atomic force microscopy (AFM). The functionalized surface was applied to develop an SPR immunosensor for detecting human serum albumin (HSA) and the estimated detection limit (DL) was 4.27 nM. The limit value concentration can be well measured between ill and healthy conditions.

The Role of Dendritic Cells in Central Tolerance

  • Oh, Jaehak;Shin, Jeoung-Sook
    • IMMUNE NETWORK
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    • v.15 no.3
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    • pp.111-120
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    • 2015
  • Dendritic cells (DCs) play a significant role in establishing self-tolerance through their ability to present self-antigens to developing T cells in the thymus. DCs are predominantly localized in the medullary region of thymus and present a broad range of self-antigens, which include tissue-restricted antigens expressed and transferred from medullary thymic epithelial cells, circulating antigens directly captured by thymic DCs through coticomedullary junction blood vessels, and peripheral tissue antigens captured and transported by peripheral tissue DCs homing to the thymus. When antigen-presenting DCs make a high affinity interaction with antigen-specific thymocytes, this interaction drives the interacting thymocytes to death, a process often referred to as negative selection, which fundamentally blocks the self-reactive thymocytes from differentiating into mature T cells. Alternatively, the interacting thymocytes differentiate into the regulatory T (Treg) cells, a distinct T cell subset with potent immune suppressive activities. The specific mechanisms by which thymic DCs differentiate Treg cells have been proposed by several laboratories. Here, we review the literatures that elucidate the contribution of thymic DCs to negative selection and Treg cell differentiation, and discusses its potential mechanisms and future directions.

Intein-mediated expression of Trichoderma reesei Cellobiohydrolase I Cellulose Binding Domain in E. coli (Intein을 이용한 대장균에서의 Trichoderma reesei 유래의 Cellobiohydrolase I 섬유소 결합 도메인의 발현)

  • Choi, Shin-Geon
    • Journal of Industrial Technology
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    • v.36
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    • pp.33-37
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    • 2016
  • Cellulose binding domains (CBDs) of cellulases are thought to assist in the hydrolysis of insoluble crystalline cellulose. To gain sufficient amount of CBDs, the self-cleavable intein tag was used for expression and purification of Trichoderma reesei cellobiohydrolase I CBD in E. coli. Synthetic CBD genes, CBD or linker-CBD were cloned into expression vector pTYB11. Recombinant CBDs were successfully purified by intein mediated purification with an affinity chitin-binding domain. The final yields of recombinant CBD and linker-CBD were 3.2 mg/L and 1.4 mg/L, respectively. The functional bindings of recombinant CBDs were confirmed by Avicel binding experiments. The simple and easy purification method using self-cleavable intein tag can be further used in pretreatment of crystalline cellulose or characterization of engineered CBDs.

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An effects on social maturity in satisfaction with the academic major of high school students majoring in culinary arts (호텔요리전공 특성화고등학생들의 사회적 학습 성취도가 요리전공만족도에 미치는 영향)

  • Park, jeong-joon
    • Journal of the Korea Convergence Society
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    • v.9 no.2
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    • pp.191-200
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    • 2018
  • In this study, the relationship between selective attribute and socio-maturity was found to be statistically significant in cooking major of cooking high school students. The selective attribute of cooking major influenced the affinity of members and the selective factor of cooking major was social Affecting the management of self - esteem and self - efficacy. The selection process of cooking major affects the superior adaptability of school life. The results of this study are as follows. First, the relationship between the social maturity and the satisfaction of the majors of the cooking major students was statistically significant in the hypothesis of the model, and the member relational affinity and social intimacy influenced the participation of the curriculum model And social maturity influenced social intimacy.

Fabrication of Nanostructured Films of Block Copolymers for Nanolithographical Masks (나노리소그래피 마스크용 블록공중합체 나노구조 필름의 제조)

  • Park Dae-Ho;Sohn Byeong-Hyeok;Jung Jin Chul;Zin Wang-Cheol
    • Journal of the Microelectronics and Packaging Society
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    • v.12 no.2 s.35
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    • pp.181-186
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    • 2005
  • We fabricated thin films of polystyrene-block-poly(methyl methacrylate)(PS -b-PMMA) on the self-assembled monolayers(SAM) of 3-(p-methoxyphenyl)propyltrichlorosilane(MPTS) on silicon wafers. Cylindrical nanodomains of PMMA or PS were oriented perpendicular to the surface of silicon wafers due to the neutral affinity of the SAM to PS and PMMA blocks. By selective removal of the PMMA block with UV irradiation and washing, nanoporous films and nanorod assemblies were produced. The nanoporous film can be used for a nanolithographical mask.

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A CPW-Fed Self-Affine Cross Shape Fractal Antenna (자기 아파인 프랙탈 구조를 이용한 CPW 급전 크로스 안테나)

  • Kim Tae-Hwan;Lee Jae-Wook;Cho Choon-Sik;Lee Yun-Hyun
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.16 no.9 s.100
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    • pp.949-956
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    • 2005
  • In this paper, a new CPW-fed cross shape fractal antenna having a self-affinity is presented. This novel configuration, which has anisotropic scaling symmetry, makes smaller profile characteristic compared to the fractal antenna using a self-similarity. Increase of the iteration coefficient, which leads to decrease of the fundamental resonant frequency, shows a good impedance matching condition and multi-band characteristics due to new surface current paths. The radiation patterns are similar to those of monopole antennas. In the K3 stage of iteration, the proposed antenna shows a measured maximum gain 2.27 dBi at 940 MHz. A commercially available software based on the FDTD algorithm has been used to obtain the predicted results. In addition, an RT/Duroid 5880 substrate has been employed for the experimental results.