• 한국미생물·생명공학회지
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• 제36권4호
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• pp.307-313
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• 2008

Bacillus sp. endoxylanase 유전자(xynB, 642 bp)의 효모 표면발현계 pCTXYN(6.8 kb)를 구축하고 Saccharomyces cerevisiae EBY100에 형질전환시켜 형질전환체 EBY100/pCTXYN를 얻었다. 형질전환체들을 xylan이 포함된 YPDG 배지에서 배양 후 활성염색을 통하여 고찰성의 형질전환체를 최종 선별하였다. 갈락토스 배지에서 자란 효모 형질전환체로부터 xynB는 성공적으로 표면발현되었고, xylan으로 부터 xylooligosaccharides를 효율적으로 생성함도 화인하였다. Endoxylanase 활성은 세포분획에서만 검출되었고 배양 48시간에 최종 1.9 unit/mL의 활성을 보였다. Xylooligosaccharides 생산을 위한 치적 반응 조건으로, 기질과 농도는 oat spelt xylan 6%, 효모 생촉매 농도는 5 unit/mL, 반응온도는 $50^{\circ}C$, 반응시간은 $2{\sim}4$시간이었다 효모 생촉매를 oat spelt xylan과 corncob xylan에 처리한 결과, xylotriose가 주성분이었다.

• Journal of Microbiology and Biotechnology
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• 제26권2호
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• pp.421-431
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• 2016

Recently, poly-γ-glutamic acid synthetase A (pgsA) has been applied to display exogenous proteins on the surface of Lactobacillus casei or Lactococcus lactis, which results in a surface-displayed component of bacteria. However, the ability of carrying genes encoded by plasmids and the expression efficiency of recombinant bacteria can be somewhat affected by the longer gene length of pgsA (1,143 bp); therefore, a truncated gene, pgsA, was generated based on the characteristics of pgsA by computational analysis. Using murine IL-10 as an exogenous gene, recombinant Lactobacillus plantarum was constructed and the capacity of the surface-displayed protein and functional differences between exogenous proteins expressed by these strains were evaluated. Surface expression of IL-10 on both recombinant bacteria with anchorins and the higher expression levels in L. plantarum-pgsA'-IL-10 were confirmed by western blot assay. Most importantly, up-regulation of IL-1β, IL-6, TNF-α, IFN-γ, and the nuclear transcription factor NF-κB p65 in RAW264.7 cells after stimulation with Poly(I:C) or LPS was exacerbated after co-culture with L. plantarum-pgsA. By contrast, IL-10 expressed by these recombinant strains could reduce these factors, and the expression of these factors was associated with recombinant strains that expressed anchorin (especially in L. plantarum-pgsA'-IL-10) and was significantly lower compared with the anchorin-free strains. These findings indicated that exogenous proteins could be successfully displayed on the surface of L. plantarum by pgsA or pgsA', and the expression of recombinant bacteria with pgsA' was superior compared with bacteria with pgsA.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.466-466
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• 2003

• Journal of Electrical Engineering and Technology
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• 제12권3호
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• pp.1314-1319
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• 2017

A handheld sheet resistance meter that can easily and quickly measure the sheet resistance of indium tin oxide films was developed. The dual-configuration four-point probe method was adopted for this instrument, which measured sheet resistance in the range from $0.26{\Omega}/sq$. to $2.6k{\Omega}/sq$. with 0.3 % ~ 0.5 % uncertainty. The screen of the instrument displayed the sheet resistance when the probe was in contact with the sample surface and the value continued to be displayed during the probe contact. Even after separating the probe from the surface, the value was still displayed on the screen and could be read easily. A feature of the instrument was the use of the dual-configuration technique to reduce edge effects markedly compared with the single-configuration technique and its ease of operation without applying correction factors for sample size and thickness.

• Journal of Microbiology and Biotechnology
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• 제24권3호
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• pp.408-420
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• 2014

Yeast surface-displayed antibody libraries provide an efficient and quantitative screening resource for given antigens, but suffer from typically modest library sizes owing to low yeast transformation efficiency. Yeast mating is an attractive method for overcoming the limit of yeast transformation to construct a large, combinatorial antibody library, but the optimal conditions have not been reported. Here, we report a large synthetic human Fab (antigen binding fragment) yeast surface-displayed library generated by stepwise optimization of yeast mating conditions. We first constructed HC (heavy chain) and LC (light chain) libraries, where all of the six CDRs (complementarity-determining regions) of the variable domains were diversified mimicking the human germline antibody repertoires by degenerate codons, onto single frameworks of VH3-23 and $V{\kappa}1$-16 germline sequences, in two haploid cells of opposite mating types. Yeast mating conditions were optimized in the order of cell density, media pH, and cell growth phase, yielding a mating efficiency of ~58% between the two haploid cells carrying HC and LC libraries. We constructed two combinatorial Fab libraries with CDR-H3 of 9 or 11 residues in length with colony diversities of more than $10^9$ by one round of yeast mating between the two haploid HC and LC libraries, with modest diversity sizes of ${\sim}10^7$. The synthetic human Fab yeast-displayed libraries exhibited relative amino acid compositions in each position of the six CDRs that were very similar to those of the designed repertoires, suggesting that they are a promising source for human Fab antibody screening.

• 한국표면공학회:학술대회논문집
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• 한국표면공학회 2011년도 춘계학술대회 및 Fine pattern PCB 표면 처리 기술 워크샵
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• pp.75-76
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• 2011

Hot-rolled steel plates of SPHC and SS400 were oxidized at 600, 750 and $900^{\circ}C$ for 2hr in air. With an increase in oxidation temperature, their oxidation rates increased, being accompanied with formation of pores and cracks in the thickened oxide scales that were non-adherent. The SPHC steels that were manufactured by either the arc furnace or the blast furnace displayed similar oxidation rates, indicating that their oxidation rates were insensitive to the manufacturing processes. The medium-carbon SS400 steel displayed a little faster oxidation rates than the low-carbon SS400 steel, indicating that the carbon content did not significantly influence the oxidation rates.

• 구강회복응용과학지
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• 제22권4호
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• pp.349-365
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• 2006

State of problem : A number of investigation about increase of surface area via various surface treatments and modification of surface constituent have been carried out. Purpose : The surface characteristics and stability of implants treated with anodic oxidation, fluoride ion incorporation, and groups treated with both methods were evaluated. Material and method : Specimens were divided into six groups, group 1 was the control group with machined surface implants, groups 2 and 3 were anodic oxidized implants (group 2 was treated with 1M $H_2SO_4$ and 185V, group 3 was treated with 0.25M $H_2SO_4$ and $H_3PO_4$ and 300V). Groups 4, 5 and 6 were treated with fluoride. Group 4 was machined implants treated with 0.1% HF, and groups 5 and 6 were groups 2 and 3 treated with 10% NaF respectively. Using variable methods, implant surface characteristics were observed, and the implant stability was evaluated on rabbit tibia at 0, 4, 8 and 12 weeks. Result : 1. In comparison of the surface characteristics of anodic oxidized groups, group 2 displayed delicate and uniform oxidation layer with small pore size containing Ti, C, O and showed mainly rutile, but group 3 displayed large pore size and irregular oxidation layer with many crators. 2. In comparison of the surface characteristics of fluoride treated groups 4, 5, 6 and non-fluoride treated groups 1, 2, 3, the configurations were similar but the fluoride treated groups displayed rougher surfaces and composition analysis revealed fluoride in groups 4, 5, 6. 3. The fluoride incorporated anodic oxidized groups showed the highest resonance frequency values and removal torque values, and the values decreased in the order of anodic oxidized groups, fluoride treated group, control group. 4. According to implant stability tests, group 2 and 3 showed significantly higher values than the control group (P<.05). The fluoride treated groups showed relatively higher values than the non fluoride treated groups and there were significant difference between group 4 and group 1 (P<.05). Conclusion : From the results above, it can be considered that the anodic oxidation method is an effective method to increase initial bone stability and osseointegration and fluoride containing implant surfaces enhance new bone formation. Implants containing both of these methods should increase osseointegration, and reduce the healing period.

• 대한의생명과학회지
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• 제18권2호
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• pp.160-164
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• 2012

The baculovirus/insect cell expression system is of great value for the large-scale production of normal and mutant mammalian passive glucose-transport proteins heterologously for structural and functional studies. In most mammalian cells that express HepG2, this transporter isoform is predominantly located at the cell surface. However, it had been reported that heterologous expression of other membrane proteins using the baculovirus system induced highly vacuolated cytoplasmic membranes. Therefore, how a cell responds to the synthesis of large amounts of a glycoprotein could be an interesting area for investigation. In order to examine the subcellular location of the human HepG2 transport proteins when expressed in insect cells, immunofluorescence studies were carried out. Insect cells were infected with the recombinant baculovirus AcNPVHIS-GT or with wild-type virus at a MOI of 5, or were not exposed to viral infection. A high level of fluorescence displayed in cells infected with the recombinant virus indicated that transporters are expressed abundantly and present on the surface of infected Sf21 cells. The evidence for the specificity of the immunostaining was strengthened by the negative results shown in the negative controls. Distribution of the transporter protein expressed in insect cells was further revealed by making a series of optical sections through an AcNPVHIS-GT-infected cell using a confocal microscope, which permits optical sectioning of cell sample. These sections displayed intense cytoplasmic immunofluorecence surrounding the region occupied by the enlarged nucleus, indicating that the expressed protein was present not only at the cell surface but also throughout the cytoplasmic membranous structures.

• 한국광학회지
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• 제8권2호
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• pp.165-168
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• 1997

780 nm의 반도체 레이저, compact disk의 광 pick-up용 actuator, 그리고 4분할 photodiode를 이용하여 scanning confocal microscope를 구성하여 시료면의 높이와 재질의 차이를 측정하였다. 4분할 photodiode에 검출되는 오차 신호를 이용하여 렌즈가 장착되어 있는 actuator를 움직이면서 시료면에 레이저 광속의 초점이 항상 위치하도록 하였으며, 이 때 actuator에 흐르고 있는 전류를 시료면의 높이로 나타내어 3차원 영상으로 표현하였다. 또한 재질의 차이는 4분할 photodiode에 검출되는 합산 신호를 이용하여 컬러 프린터에 나타나는 3차원 영상의 색을 다르게 나타내었다. 전체적인 크기도 30 mm * 20 mm * 20 mm 로서 작고 간단하며 scan 영역은 최대 1.6 mm * 1.6 mm이다. 반사광의 세기를 이용한 scanning confocal microscope의 영상과 4분할 photodiode에 검출되는 오차 신호를 적분하는 방식을 이용한 scanning confocal microscope의 영상을 구하여 그 차이를 비교하였다.

• 대한전기학회:학술대회논문집
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• 대한전기학회 1998년도 하계학술대회 논문집 E
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• pp.1824-1827
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• 1998

A Laser Doppler Vibrometer(LDV) was developed using He-Ne laser as a light source. The heterodyne method was employed and its output signal was digitally processed with a $\mu$-processor and the result was displayed with LCD. The frequency shifted object beam(40 MHz) by a Bragg cell was focused on the surface of the moving target and the Doppler shifted reflected beam was recombined with reference beam at the fast photodetector to produce frequency modulated signal centered at 40 MHz. The signal from the detector was amplified and downconverted to intermediate frequency centered at 1 MHz after mixing process. The voltage output that was proportional to the velocity of the moving surface was obtained using PLL. With the same method, the fringe pattern signal of the moving surface is obtained. This fringe pattern signal is converted to TTL signal with ZCD(zero-crossing detector) and then counted to calculate the displacement due to the vibration, which is displayed with LCD. This LDV can be used to measure the resonant frequency of the electric equipments such as circuit breakers and transformers, of which resonant frequencies are changed when they are damaged.