• Title/Summary/Keyword: T/T probe

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Development of a real-time PCR method for detection and quantification of the parasitic protozoan Perkinsus olseni

  • Gajamange, Dinesh;Yoon, Jong-Man;Park, Kyung-Il
    • The Korean Journal of Malacology
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    • v.27 no.4
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    • pp.387-393
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    • 2011
  • The objective of this study was to develop a real-time PCR method for the rapid detection and quantification of the protozoan pathogen Perkinsus olseni using a TaqMan probe. For the standard, genomic DNA was extracted from $10^5$ in vitro-cultured P. olseni trophozoites, and then 10-fold serial dilutions to the level of a single cell were prepared. To test the reliability of the technique, triplicates of genomic DNA were extracted from $5{\times}10^4$ cells and 10-fold serial dilutions to the level of 5 cells were prepared. The standards and samples were analyzed in duplicate using an $Exicycler^{TM}$ 96 real-time quantitative thermal block. For quantification, the threshold cycle ($C_T$) values of samples were compared with those obtained from standard dilutions. There was a strong linear relationship between the $C_T$ value and the log concentration of cells in the standard ($r^2$ = 0.996). Detection of DNA at a concentration as low as the equivalent of a single cell showed that the assay was sensitive enough to detect a single cell of P. olseni. The estimated number of P. olseni cells was similar to the original cell concentrations, indicating the reliability of P. olseni quantification by real-time PCR. Accordingly, the designed primers and probe may be used for the rapid detection and quantification of P. olseni from clam tissue, environmental water, and sediment samples.

T-OLED의 반사전극으로 사용하기 위한 Ag 박막 표면의 UV에 의한 산화 및 KPFM을 이용한 표면 전위 측정

  • Kim, Seong-Jun;Kim, Su-In;Kim, Dong-Uk;Kim, Ju-Yeon;Lee, Eun-Hyeok;Sin, Dong-Hun;Lee, Chang-U
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.182.1-182.1
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    • 2013
  • Silver (Ag)는 높은 반사율을 가지고 있어 Top-Emission Organic Light Emitting Diode (T-OLED)의 반사전극으로 사용하기 적합하지만 일함수가 낮은 단점 (4.3 eV)을 가지고 있다. 이런 낮은 일함수를 증가시키기 위하여 Ag 박막 표면을 산화시켜 일함수를 증가시키기 위한 연구가 진행중에 있으며, 이 연구에서는 UV로 $O_3$을 발생시켜 Ag 박막 표면을 산화시키기 위한 연구를 진행하였다. 특히, Ag 박막 표면의 일함수 변화를 측정하기 위하여 SPM (Scanning Probe Microscopy)의 KPFM (Kelvin Probe Force Microscopy) mode를 적용하여 nano 영역에서의 일함수 변화를 surface potential로 측정하여 UV 표면 산화에 의한 표면 일함수 형상을 확인하였다. Ag 박막은 rf magnetron sputter를 사용하여, Si 기판위에 300nm 두께로 증착시켰다. 이후 $O_3$ 발생되는 UV 램프로 Ag 박막 표면 30초 간격으로 최대 5분간 산화시켰으며, 이후 KPFM mode를 사용하여 산화 시간에 따른 Ag 박막 표면의 potential 변화를 측정하였다. 0~3분간 산화된 Ag 박막 표면의 potential은 약 6 mV로 일정하였으나 3분 이후 최대 110 mV까지 급격하게 변화하는 것을 확인할 수 있었다. Ag 박막 표면의 RMS roughness는 UV 산화처리 전0.7 nm였으나, potential이 급격하게 증가하는 시점인 3분 이후 2.83 nm로 약 400% 이상 증가하였다. 이를 통해 $O_3$ 발생 UV 램프로 산화된 Ag 박막의 표면 물성은 처리 시간에 따라 급격히 변하는 것을 확인하였다.

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Gluconacetobacter persimmonis sp. nov., Isolated from Korean Traditional Persimmon Vinegar

  • Yeo, Soo-Hwan;Lee, Oh-Seuk;Lee, In-Seon;Kim, Hyun-Soo;Yu, Tae-Shick;Jeong, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.276-283
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    • 2004
  • Screening was performed to isolate cellulose-producing microorganisms from the Korean traditional fermented persimmon vinegar. The resulting strain, KJ $145^{T}$, was then taxonomically investigated by phenotypic characterization, particularly chemotaxonomic, and by phylogenetic inference based on a 16S rDNA sequence analysis including other related taxa. Strain KJ $145^{T}$ was found to grow rapidly and form pale white colonies with smooth to rough surfaces on a GYC agar. Strain KJ $145^T$ also produced acetate from ethanol, and was tolerable to 10% ethanol in SM medium. In a static culture, a thick cellulose pellicle was produced, and in GYC broth, the strain grew at temperatures ranging from 28 to $40^\circ{C}$ with an optimum pH of 4.0. The genomic DNA G+C content of strain KJ $145^T$ was 61.9 mol%, and the predominant ubiquinone was Q 10 as the major quinone and Q9 as the minor quinone. The major cellular fatty acids were $C_{16:0}$ and the sum in feature 7 ($C_{18:1}$ w9c, w12t and/or w7c). A 16S rRNA-targeted oligonucleotide probe specific for strain KJ $145^T$was constructed, and the phylogenetic position of the new species was derived from a 16S rDNA-based tree. When comparing the 16S rDNA nucleotide sequences, strain KJ $145^T$ was found to be most closely related to G. hansenii LMG $1527^T$ (99.2%), although KJ $145^T$ was still distinct from G. hansenii LMG $l527^T$ and G. xylinus LMG $1515^T$ in certain phenotypic characteristics. Therefore, on the basis of 16S rDNA sequences and taxonomic characteristics, it is proposed that strain KJ $145^T$ should be placed in the genus Gluconacetobacter as a new species, Gluconacetobacter persimmonis sp. nov., under the type-strain KJ $145^T$ (=KCTC =$10175BP^T$=KCCM=$10354^T$).

An improvement of real-time polymerase chain reaction system based on probe modification is required for accurate detection of African swine fever virus in clinical samples in Vietnam

  • Tran, Ha Thi Thanh;Dang, Anh Kieu;Ly, Duc Viet;Vu, Hao Thi;Hoang, Tuan Van;Nguyen, Chinh Thi;Chu, Nhu Thi;Nguyen, Vinh The;Nguyen, Huyen Thi;Truong, Anh Duc;Pham, Ngoc Thi;Dang, Hoang Vu
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.10
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    • pp.1683-1690
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    • 2020
  • Objective: The rapid and reliable detection of the African swine fever virus (ASFV) plays an important role in emergency control and preventive measures of ASF. Some methods have been recommended by FAO/OIE to detect ASFV in clinical samples, including realtime polymerase chain reaction (PCR). However, mismatches in primer and probe binding regions may cause a false-negative result. Here, a slight modification in probe sequence has been conducted to improve the qualification of real-time PCR based on World Organization for Animal Health (OIE) protocol for accurate detection of ASFV in field samples in Vietnam. Methods: Seven positive confirmed samples (four samples have no mismatch, and three samples contained one mutation in probe binding sites) were used to establish novel real-time PCR with slightly modified probe (Y = C or T) in comparison with original probe recommended by OIE. Results: Both real-time PCRs using the OIE-recommended probe and novel modified probe can detect ASFV in clinical samples without mismatch in probe binding site. A high correlation of cycle quantification (Cq) values was observed in which Cq values obtained from both probes arranged from 22 to 25, suggesting that modified probe sequence does not impede the qualification of real-time PCR to detect ASFV in clinical samples. However, the samples with one mutation in probe binding sites were ASFV negative with OIE recommended probe but positive with our modified probe (Cq value ranked between 33.12-35.78). Conclusion: We demonstrated for the first time that a mismatch in probe binding regions caused a false negative result by OIE recommended real-time PCR, and a slightly modified probe is required to enhance the sensitivity and obtain an ASF accurate diagnosis in field samples in Vietnam.

Expression of Antisense Polygalacturonase Gene in Transgenic Tomato (형질전환 토마토에서 Antisense Polygalacturonase 유전자의 발현)

  • 김영미;김용환;이성갑;임명호;송경수
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.6
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    • pp.351-355
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    • 1995
  • A truncated Polygalacturonase (PG) cDNA was fused in reverse orientation to the CaMV 35S promoter of the binary vector pCA643, and introduced into tomato cells by Agrobaderium - mediated transformation. Transformed cells were selected using kanamycin as select agent then regenerated into plants. After selfed, one transgenic line (T9), was germinated and grown on MS medium containing 1 mg/mL of kanamycin Genomic Southern analysis of a T9 progeny with labelled PG2 cDNA probe showed a single antisense PC fragment as well as the endogenous PG2 gene, suggesting that PC antisense gene was integrated into tomato genome. Northern blot analysis demonstrated that the antisense RNA was produced from the transgene at much tiger level than the endogenous PG2 gene. Polygalacturonase activity analysis of the fruit from transgenic plants demonstrated that the antisense transgene expression caused 4 to 60% reduction of endogenous PG activity.

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RADIO EMISSION FROM AKARI GALAXIES

  • Pepiak, A.;Solarz, A.;Pollo, A.;Takeuchi, T.T.;Jurusik, W.;AKARI Team, AKARI Team
    • Publications of The Korean Astronomical Society
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    • v.27 no.4
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    • pp.339-341
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    • 2012
  • It is a long known fact that there exists a tight correlation between far-infrared and radio emission both for galaxies hosting active galactic nuclei and for star forming galaxies. We probe the radio - infrared correlation for a sample of extragalactic sources constructed by the cross-correlation of the AKARI/IRC All-Sky Survey Point Source Catalogue, the AKARI/FIS All-Sky Survey Bright Source Catalogue, and the NRAO VLA Sky Survey. Additionally, all objects of our sample were identified as galaxies in NED and SIMBAD databases, and a part of them is known to host active galactic nuclei (AGNs). After remeasuring all the fluxes, in order to avoid small aperture effects, we compare the ratio of radio to infrared emission from different types of extragalactic sources, and discuss the FIR/radio correlation as seen by AKARI and make a comparison to the previous results obtained thanks to IRAS.

Molecular Cloning of Antagonistic Genes in Pseudomonas maItophiliQ B-14 (토양병해 길항성 Pseudomonas maltophilia B-14의 길항유전자탐색)

  • 구본성;서영우;윤상홍;박경수;은무영;김용환;오상우;류진창;은무영
    • Microbiology and Biotechnology Letters
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    • v.20 no.6
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    • pp.619-624
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    • 1992
  • Tn5 lac 삽입으로 채소입고병원균에 길항력이 약화된 T-67 및 고추역병균과 참깨역병균에 길항력이 약화된 T-81의 Tn5 lac 유전자 일부와 오른쪽 말단에 있는 길항관련 유전자의 flanking sequence가 cloning된 pAG67 및 pAG81 clone을 선발하였고, pAG67 및 pAG81 clone된 길항관련 유전자의 flanking sequence를 야생 길항균 Pseudomonas maltophilia B-14의 DNA를 probe로 사용하여 Southern hybridization으로 확인하였으며, 제한효소 지도를 작성하여 8Kb 및 4Kb 크기의 flanking sequence가 cloning되었음을 확인하였다. pAG6 및 pAG81의 flanking sequence를 EcoRi-BglII와 EcoRI-MpaI으로 분리하여 유전자 은행으로부터 길항관련 유전자가 cloning된 cosmid clone 7개주를 선발하였다.

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Activated Physical Properties at Air-Polymer Interface

  • Kajiyama, Tisato;Tanaka, Keiji
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.5-6
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    • 2006
  • Molecular motion at the surface of monodisperse polystyrene (PS) films with various chain end groups was studied by scanning probe microscopy. Surface glass transition temperature ($T_{g^s}$) of the PS films was much lower than the corresponding bulk value. And, the magnitude of $T_{g^s}$ was strongly dependent on chain end chemistry. This result can be explained in terms of the chain end concentration at the surface. Time-temperature superposition principle was applied to rheological analysis at the surface. The apparent activation energy of the surface ${\alpha}_{a}$-relaxation process was approximately a half of that for the bulk sample. This result clearly indicates that the cooperativity for the surface segmental motion was reduced in comparison with that in the bulk region.

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Fine YBaCuO Powder Prepared by Sol-gel Method

  • Soh, Deawha;Li, Fenghua;Wang, Jue;Fan, Zhanguo
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2002.07a
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    • pp.470-473
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    • 2002
  • In order to get proper original suspension for the electrophoresis process of making YBaCuO films, YBaCuO superconductor powder was prepared with the Sol-gel method. The composition of the powder was analyzed by X-ray diffraction, which was identified as YBa$_2$Cu$_3$O$\sub$7-x/(Yl23) phase. The form of YBaCuO single particle was showed to be spherical by scanning electronic microscope and its size was among 0.2-1 $\mu\textrm{m}$. The critical transition temperature (T$\sub$c/) and the critical current density (J$\sub$c/) were measured with the four-probe, method. The T$\sub$c/ was about 91 K, and the J$\sub$c/ was 5-30 A/$\textrm{cm}^2$.

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A Voltage Sensor for measurement of Very Fast Transients in GIS (GIS에 있어서의 과도과전압(過渡過電壓) 측정용(測定用) 센서)

  • Lee, B.H.;Paek, Y.H.;Kawamura, T.;Nishimura, T.;Ishii, M.
    • Proceedings of the KIEE Conference
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    • 1990.07a
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    • pp.297-300
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    • 1990
  • The measurement of very fast transients generated by disconnecting switches in gas-insulated switchgear(GIS) have to deal with the problems such as reliability, interference pich-up, optimal design of high voitage equipments. This paper presents a new developed voltage sensor to measure the very fast transients, the basic theory of the measuring method, the design, structure of planar capacitive voltage probe are described. Finally the examples of modelling tests on an actual site GIS are discussed.

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