• 제목/요약/키워드: surface-displayed

검색결과 453건 처리시간 0.031초

Optimization of Yeast Surface-Displayed cDNA Library Screening for Low Abundance Targets

  • Kim, Juhyung;Kim, Hyung Kyu;Jang, Hye Jeong;Kim, Eunkyung;Kim, Moon Kyu
    • Journal of Microbiology and Biotechnology
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    • 제25권4호
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    • pp.547-553
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    • 2015
  • The yeast surface-displayed cDNA library has been used to identify unknown antigens. However, when unknown target antigens show moderate-to-low abundance, some modifications are needed in the screening process. In this study, a directional random-primed cDNA library was used to increase the number of candidates for the unknown antigen. To avoid the loss of target yeast clones that express proteins at a low frequency in the cDNA library, a comprehensive monitoring system based on magnetic-activated cell sorting, fluorescence-activated cell sorting, and immunofluorescence was established, and a small number of target yeast cells was successfully enriched. These results showed that our optimized method has potential application for identifying rare unknown antigens of the human monoclonal antibody.

Comparison of the Organophosphorus Hydrolase Surface Display Using InaVN and Lpp-OmpA Systems in Escherichia coli

  • Karami, Ali;Latifi, Ali Mohamad;Khodi, Samaneh
    • Journal of Microbiology and Biotechnology
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    • 제24권3호
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    • pp.379-385
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    • 2014
  • The purpose of this study was to compare the ability of an engineered Escherichia coli to degrade chlorpyrifos (Cp) using an organophosphorus hydrolase enzyme, encoded in both Flavobacterium sp. ATCC 27551 or Pseudomonas diminuta, by employing the Lpp-OmpA chimera and the N-terminal domain of the ice nucleation protein as anchoring motifs. Tracing of the expression location of the recombinant protein using SDS-PAGE showed the presentation of OPH by both anchors on the outer membrane. This is the first report on the presentation of OPH on the cell surface by Lpp-OmpA under the control of the T7 promoter. The results showed cell growth in the presence of Cp as the sole source of energy, without growth inhibition, and with higher whole-cell activity for both cells harboring plasmids pENVO and pELMO, at approximately 10,342.85 and 10,857.14 U/mg, respectively. Noticeably, the protein displayed by pELMO was lower than the protein displayed by pENVO. It can be concluded that Lpp-OmpA can display less protein, but more functional OPH protein. These results highlight the high potential, of both engineered bacteria, for use in the bioremediation of pesticide-contaminated sources in the environment.

Fe합금의 내 산화성과 황화성에 미치는 Al과 Cr 함량의 영향 (Effect of Al and Cr contents on the High Temperature Oxidation- and Sulfidation-resistance of Fe Alloys)

  • 김슬기;이동복
    • 한국표면공학회지
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    • 제45권2호
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    • pp.61-69
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    • 2012
  • Alloys of Fe-(5, 10, 15)Al and Fe-(10, 20, 30, 40)Cr were corroded at 700 and $800^{\circ}C$ for 70 hr in either atmospheric air or 1 atm of Ar+$1%SO_2$ gases. In these atmospheres, Fe-5Al and Fe-10Cr alloys displayed poor corrosion resistance. In atmospheric air, Fe-5Al alloys formed oxide nodules, while Fe-10Cr alloys formed thick scales and internal oxides. In Ar+$1%SO_2$ gases, Fe-5Al and Fe-10Cr alloys formed thick, nonadherent bi-layered scales, which grew primarily by the outward diffusion of Fe ions and inward diffusion of oxygen and sulfur ions. By contrast, in atmospheric air and Ar+$1%SO_2$ gases, Fe-(10, 15)Al and Fe-(20, 30, 40)Cr alloys displayed good corrosion resistance by forming $Al_2O_3$ and $Cr_2O_3$ layers on the surface, respectively.

효모 표층 Arylsulfatase에 의해 제조된 Agarose의 특성 (Characterization of Agarose Produced by Yeast Cell Surface Displayed-Arylsulfatase)

  • 조은수;김정환;김연희;남수완
    • 한국미생물·생명공학회지
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    • 제38권4호
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    • pp.428-433
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    • 2010
  • Agar로부터 agarose 제조 시 유기용매를 이용해서 황을 제거하는 방법이 일반적으로 많이 사용되어진다. 하지만 agar의 황을 가수 분해하는 효소를 사용할 경우 agarose 제조 시공정과정을 획기적으로 간소화할 수 있다. 따라서 arylsulfatase로 agaropectin에서 황 제거를 통해 agarose로 바꾸는 공정은 간단하고 높은 수율을 얻을 수 있기 때문에 agarose 생산에 효율적으로 적용할 수 있다. 본 연구에서는 arylsulfatase를 세포표면에 발현하는 효모생촉매를 이용하여 제조한 agarose의 황 함량과 gel 강도를 측정하였다. 처리한 효모(효)소의 농도가 증가할수록 증가된 탈황 반응에 의해 황함량이 줄어들었고, 특히 35 unit/mL의 효소 농도로 처리하였을 때 황 함량은 0.2%까지 감소하는 것을 확인할 수 있었다. 황 함량을 가장 낮출 수 있는 최적 조건은 0.6% agar(Junsei) 용액에 효모 표층 arylsulfatase 35 unit/mL로 처리하고 $40^{\circ}C$에서 3시간 반응시켰을 때 였다. 또한 1.0% DNA 전기 영동용 agarose의 gel 강도는 효모 표층 arylsulfatase 처리로 제조된 agarose의 경우 $559.8{\pm}0.12$로 상업적 agarose의 gel 강도($880.6{\pm}0.15\;g/cm^2$와) 보다는 낮았다. 따라서 효모 S. cerevisiae의 세포 표면에서 발현된 재조합 arylsulfatase 효소를 이용하여 agar로부터 전기영동용 agarose의 생산 공정에 적용 가능함을 알 수 있었다.

Removal of Chromium (VI) by Escherichia coli Cells Expressing Cytoplasmic or Surface-Displayed ChrB: a Comparative Study

  • Zhou, Xiaofeng;Li, Jianghui;Wang, Weilong;Yang, Fan;Fan, Bingqian;Zhang, Chenlu;Ren, Xiaojun;Liang, Feng;Cheng, Rong;Jiang, Fengying;Zhou, Huaibin;Yang, Juanjuan;Tan, Guoqiang;Lyu, Jianxin;Wang, Wu
    • Journal of Microbiology and Biotechnology
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    • 제30권7호
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    • pp.996-1004
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    • 2020
  • Various genetically engineered microorganisms have been developed for the removal of heavy metal contaminants. Metal biosorption by whole-cell biosorbents can be enhanced by overproduction of metal-binding proteins/peptides in the cytoplasm or on the cell surface. However, few studies have compared the biosorption capacity of whole cells expressing intracellular or surface-displayed metal-adsorbing proteins. In this study, several constructs were prepared for expressing intracellular and surface-displayed Ochrobactrum tritici 5bvl1 ChrB in Escherichia coli BL21(DE3) cells. E. coli cells expressing surface-displayed ChrB removed more Cr(VI) from aqueous solutions than cells with cytoplasmic ChrB under the same conditions. However, intracellular ChrB was less susceptible to variation in extracellular conditions (pH and ionic strength), and more effectively removed Cr(VI) from industrial wastewater than the surface-displayed ChrB at low pH (<3). An adsorption-desorption experiment demonstrated that compared with intracellular accumulation, cell-surface adsorption is reversible, which allows easy desorption of the adsorbed metal ions and regeneration of the bioadsorbent. In addition, an intrinsic ChrB protein fluorescence assay suggested that pH and salinity may influence the Cr(VI) adsorption capacity of ChrB-expressing E. coli cells by modulating the ChrB protein conformation. Although the characteristics of ChrB may not be universal for all metal-binding proteins, our study provides new insights into different engineering strategies for whole-cell biosorbents for removing heavy metals from industrial effluents.

평면연삭(平面硏削)에서 주파수(周波數) 분석(分析)에 의한 표면(表面) 거칠기 해석(解釋) (A Study on the Surface Roughness in Grinding Process by Frequency Analysis)

  • 김경년;정재천;김기선
    • 비파괴검사학회지
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    • 제11권2호
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    • pp.32-41
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    • 1991
  • This paper is to analyze the surface roughness in wet and dry surface grinding in which working conditions are table feed, cross feed and down feed. FFT analyzer is used to identify the surface roughness and the data of surface roughness are stored and displayed using an interfaced computer. In results, components affecting surface roughness exist within 200Hz with respect the conditions and methods of grinding operation. Also, with increase of table feed, cross feed and down feed, the value of surface roughness increases. Of them, cross feed has first influence on surface roughness. In dry, preciser surface can be obtained than one in wet.

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전해도금에 의한 Ni-C 복합층의 내식성 및 표면 전기저항 (Corrosion and Surface Resistance of Ni-C Composite by Electrodeposition)

  • 박제식;이성형;정구진;이철경
    • 한국재료학회지
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    • 제21권5호
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    • pp.288-294
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    • 2011
  • Simultaneous Ni and C codeposition by electrolysis was investigated with the aim of obtaining better corrosion resistivity and surface conductivity of a metallic bipolar plate for application in fuel cells and redox flow batteries. The carbon content in the Ni-C composite plate fell in a range of 9.2~26.2 at.% as the amount of carbon in the Ni Watt bath and the roughness of the composite were increased. The Ni-C composite with more than 21.6 at.% C content did not show uniformly dispersed carbon. It also displayed micro-sized defects such as cracks and crevices, which result in pitting or crevice corrosion. The corrosion resistance of the Ni-C composite in sulfuric acid is similar with that of pure Ni. Electrochemical test results such as passivation were not satisfactory; however, the Ni-C composite still displayed less than $10^{-4}$ $A/cm^2$ passivation current density. Passivation by an anodizing technique could yield better corrosion resistance in the Ni-C composite, approaching that of pure Ni plating. Surface resistivity of pure Ni after passivation was increased by about 8% compared to pure Ni. On the other hand, the surface resistivity of the Ni-C composite with 13 at.% C content was increased by only 1%. It can be confirmed that the metal plate electrodeposited Ni-C composite can be applied as a bipolar plate for fuel cells and redox flow batteries.

Open 균일 B-spline 곡면을 이용한 선체 곡면 표현에 관한 연구 (A Study of Geometric Modeling for Ship Hull Forms Using Open Uniform B-spline Surface)

  • 신현경;박규원
    • 대한조선학회논문집
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    • 제28권2호
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    • pp.21-27
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    • 1991
  • 이 논문에서는 periodic 균일 knot vector 뿐만아니라 open 균일 knot vector를 사용하여 선체형상을 Bi-cubic B-spline곡면으로 수식화하는 방법을 보인다. B-spline곡면을 형성하기 위한 B-spline control vertex는 기본 함수의 pseudoinverse matrix를 사용하여 결정된다. 주어진 offset과 형성된 선체곡면을 비교한 결과 잘 일치하였다. 곡면의 순정을 검토하기 위하여 Gaussian곡률을 많은 작은 곡면조각에 대해 계산하여 흑백의 농도 차이를 이용하여 도시화하였다.

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