• 제목/요약/키워드: Tn5 cells

검색결과 73건 처리시간 0.03초

$aroP^{-}$변이가 E.coli에서 트립토판 방출에 미치는 영향 (Effects of $aroP^{-}$ mutation on the tryptophan excretion in escherichia coli)

  • 지연태;안병우;이세영
    • 미생물학회지
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    • 제23권1호
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    • pp.9-12
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    • 1985
  • 증폭된 재조합 trp operon의 발현을 위한 숙주박테리아 개발의 일환으로 숙수 E. coli에 $aroP^{-}$ 변이를 도입하였다. $aroP^{-}$ 변이의 유도에는 trans po son Tn10을 사용하였으며 P1Kc파아지를 이용하여 숙주박테리아에 형질도입하였다. General aromatic amino acid transport system이 결여된 $aroP^{-}$ 변이주는 $\beta$-thienylalanine ($(2{\times}10^{-4}M)$). p-fluor-phenylalanine ($(2{\times}10^{-4}M)$) 그리고 5-methyltryptophan에 저항성을 가졌다. $aroP^{-}$ 변이주는 $aroP^{-}$ 야생주에 비해서 〔$[^3H]$-tryptophan uptake가 상당히 감소하였다. 또한 NaN, ($(2{\times}10^{-4}M)$)를 처리하였을 때의 ($[^3H]$)-tryptophan uptake 비율은 aroP 변이주가 $aroP^{-}$야생주보다 덜 감소하였다. E. coli $trpR^{-ts}/ColE_1 -trp^+$ 균주에 aroP 형질을 도입하였을 때 트립토판 방출이 $aroP^{-}$ 야생주에 비해서 4 배나 증가하였다.

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Modulation of Human Cardiac Progenitors via Hypoxia-ERK Circuit Improves their Functional Bioactivities

  • Jung, Seok Yun;Choi, Sung Hyun;Yoo, So Young;Baek, Sang Hong;Kwon, Sang Mo
    • Biomolecules & Therapeutics
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    • 제21권3호
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    • pp.196-203
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    • 2013
  • Recent accumulating studies have reported that hypoxic preconditioning during ex vivo expansion enhanced the self-renewal or differentiation of various stem cells and provide an important strategy for the adequate modulation of oxygen in culture conditions, which might increase the functional bioactivity of these cells for cardiac regeneration. In this study, we proposed a novel priming protocol to increase the functional bioactivity of cardiac progenitor cells (CPCs) for the treatment of cardiac regeneration. Firstly, patient-derived c-$kit^+$ CPCs isolated from the atrium of human hearts by enzymatic digestion and secondly, pivotal target molecules identified their differentiation into specific cell lineages. We observed that hCPCs, in response to hypoxia, strongly activated ERK phosphorylation in ex vivo culture conditioning. Interestingly, pre-treatment with an ERK inhibitor, U0126, significantly enhanced cellular proliferation and tubular formation capacities of CPCs. Furthermore, we observed that hCPCs efficiently maintained the expression of the c-kit, a typical stem cell marker of CPCs, under both hypoxic conditioning and ERK inhibition. We also show that hCPCs, after preconditioning of both hypoxic and ERK inhibition, are capable of differentiating into smooth muscle cells (SMCs) and cardiomyocytes (CMs), but not endothelial cells (ECs), as demonstrated by the strong expression of ${\alpha}$-SMA, Nkx2.5, and cTnT, respectively. From our results, we conclude that the functional bioactivity of patient-derived hCPCs and their ability to differentiate into SMCs and CMs can be efficiently increased under specifically defined culture conditions such as short-term hypoxic preconditioning and ERK inhibition.

Expression and Characterization of Recombinant E2 Protein of Hepatitis C Virus by Insect Cell/Baculovirus Expression System

  • Han, Bong-Kwan;Lee, Bum-Yong;Min, Mi-Kyung;Jung, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제8권4호
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    • pp.361-368
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    • 1998
  • The E2 protein of HCV (hepatitis C virus) is thought to have a potential role in the development of subunit vaccines and diagnostics. To express it by the insect cell/baculovirus expression (Bacu) system, we constructed a recombinant Autographa californica nuclear polyhedrosis virus (AcIL3E2), determined the most appropriate expression conditions in terms of host cell line and culture medium, and characterized the expressed HCV E2 protein. A culture system using Trichoplusia ni BTI-TN5Bl-4 cells and SF 900IISFM medium expressed a relatively high level of HCV E2 protein. It was revealed that its glycosylation properties and subcellular localization were almost the same as the ones in the mammalian cell expression system previously reported, suggesting the recombinant HCV E2 protein derived from our Bacu system can be utilized for development of a subunit vaccine and diagnostics. Interestingly, HCV E2 protein was not degraded at all even at 43 h post-heat shock in the heat shock-induced necrotic cells, probably due to its integration into the microsomal membrane, indicating that heat shock can be employed to purify HCV E2 protein.

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Construction of Recombinant Xanthomonas campestris Strain Producing Insecticidal Protein of Bacillus thuringiensis

  • Shin, Byung-Sik;Koo, Bon-Tag;Choi, Soo-Keun;Park, Seung-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제4권4호
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    • pp.285-289
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    • 1994
  • An insecticidal crystal protein gene, cryIA(c), from Bacillus thuringiensis HD-73 was integrated into the chromosome of a xanthan-producing bacterium, Xanthomonas campestris XP92. The cryIA(c) gene expression cassette was constructed that placed the gene between the trc promoter and rrnB transcriptional terminator. The $lacl^q$ gene was also included to prevent the expression of cryIA(c) gene in X campestris cells. Southem blot analysis confirmed the integration of the cryIA(c) gene expression cassette in chromosome of X campestris XP92 transconjugant. Expression of the insecticidal crystal protein was confirmed by Western blot analysis and bioassay against the larvae of Hyphantria cunea (Lepidoptera: Arctiidae) and Plutella xylostella (Lepidoptera:Plutellidae).

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색 편광판을 이용한 3층 구조 TN LCD의 설계 (Design of a 3-stack color LCD using color polarizers)

  • 박경호;이기동;윤태훈;김재창
    • 한국광학회지
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    • 제12권5호
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    • pp.425-428
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    • 2001
  • 본 논문에서는 감색 혼법을 이용하는 3층 구조 LCD를 제시하였다. 제안된 구조는 1장의 무색 편광판과 3장의 색 편광판을 이용하여, 색 편광판만을 이용할 때 생기는 빛의 누설현상을 줄여, 명암 대비비를 향상시키는 효과를 얻었다. 그리고, 높은 색 특성을 나타내기 위해 색 편광판의 순서를 최적화 하였다.

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동해안 석호의 육수학적 조사(2): 향호 (The Limnological Survey of a Coastal Lagoon in Korea (2): Lake Hyangho)

  • 권상용;이재일;김동진;김범철;허우명
    • 생태와환경
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    • 제37권1호통권106호
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    • pp.1-11
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    • 2004
  • 향호의 육수생태학적 특성에 관한 조사를 1998년 5월부터 2002년 11월까지 겨울을 제외하고 2개월 간격으로 실시하였다. 수온, 염분, 투명도, 총질소, 총인, 및 퇴적물의 유기물량과 같은 물리 화학적인 조사와 엽록소 a 농도, 식물플랑크톤 우점종 및 밀도 변화 등과 같은 생물 조사를 병행 하였다. 향호에서 화학성층은 2 ${\sim}$ 5 m 수심에서 관측되었으며, 용존산소 농도 및 수온분포에 영향을 주는 것으로 나타났다. 염분이 높은 수층에서 용존산소의 농도가 낮았으며, 수직혼합의 제한으로 11월에는 심층에서 표층보다 약간 높은 수온 역전현상이 관측되었다. 투명도는 0.1 ${\pm}$ 0.0 ${\sim}$ 0.1 m로 매우 낮은 수준이였다. 표층에서 총인, 총질소 및 엽록소 a 농도는 각각 0.011 ${\sim}$ 0.238mgP $L^{-l}$, 0.4 ${\sim}$ 2.4 mgN $L^{-l}$ 및 0.7 ${\sim}$ 145.2 mg $m^{-3}$이였다. TN/TP 비는 대부분 30 이하였으나, 총질소와 총인농도는 높았다. 퇴적물의 입자크기는 0 ${\sim}$ 125 ${\mu}m$로 silt와 coarse silt이였다. 퇴적물의 화학적산소요구량, 총인 및 총질소는 각각 19.7 ${\sim}$ 73.3 mg$O_2\;gdw^{-1}$, 0.61 ${\sim}$ l.32 mgP $gdw^{-l}$ 및 0.64 ${\sim}$ 0.88 mgN $gdw^{-l}$의 범위이였다. 식물플랑크톤은 시기에 따라 차이는 있으나 녹조류인 Ankistrodesmus falcatus, 남조류인 Oscillatoria sp. 및 Merismopedia tennuissima 등이 우점하였다. 총 세포수는 560 ${\sim}$ 35,255 cells $mL^{-l}$의 범위로 계절적으로 끈 차이를 보였다.

Autocrine mechanism for viability enhancement of BAL eosinophils after segmental antigen challenge in allergic asthmatics.

  • Cho, Seung-Kil;Stephen P. Peters;Kim, Chang-Jong
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1996년도 춘계학술대회
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    • pp.254-254
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    • 1996
  • Eosinophils are known to be important effector cells in pathogenesis of asthma. The elucidation of mechanism by which eosinophil survival is regulated in vivo at sites of inflammation is critical tn our understanding of asthma pathogenesis. The maintenance of these cells at site of inflammation depends upon tile balance between its tendency to undergo apoptosis and tile local eosinophil-viability enhancing activity, Qualitative and quantative phenotypic differences have been observed between bronchoalveolar lavage (BAL) and peripheral blood (PB) eosinophils (EOS). We hypothesize that BAL EOS Possess altered functional feature compared to PB EOS. BAL and PB EOS were obtained from ragweed allergic asthmatics after segmental antigen challenge (SAC) at 24 hour or one week, and purified over percoll and CDl6 negative selection. Cells were cultured in duplicate in RPMI, 15% FCS and 1% penicillin/streptomycin without exogenous cytokines. Eosinophil purity and viability was >92%. BAL. EOS viability was 69${\pm}$4.4% versus 39${\pm}$1.6% for PB EOS (p<0.005) at 48 hour time point, and this difference was maintained through day 5 (32${\pm}$7.6% vs. 3.0${\pm}$ 1.4%, p<0.05), Among BAL EOS, those harvested one week after SAC appeared to have an prolonged survival compared to those harvested at 24 hour. Coculture of BAL and PB EOS resulted in significant viability enhancement than expecteed. Direct neutralization of GM-CSF activity, not IL-3 and EL-5, markedly decreased tile survival of BAL EOS in culture, and abrogated tile viability enhancing activity of their culture supernatants in a dose dependent manner. We conclude that BAL EOS activated in vivo possess enhanced viability compared to PB EOS. Mixing and neutralization experiments suggest a role for autocrine production of GM-CSF.

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이온빔 에너지에 따른 액정배향막의 전기광학적 특성연구 (Properties of liquid crystal alignment layers exposued to ion-beam irradiation enemies)

  • 오병윤;이강민;박홍규;김병용;강동훈;한진우;김영환;한정민;이상극;서대식
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2007년도 추계학술대회 논문집
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    • pp.430-430
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    • 2007
  • In general, polyimides (PIs) are used in liquid crystal displays (LCDs) as alignment layer of liquid crystals (LCs). Up to date, the rubbing alignment technique has been widely used to align liquid crystals on the PI surface, which is suitable for mass-production of LCDs because of its simple process and high productivity. However, this method has some disadvantages. Rubbed PI surfaces include the debris left by the cloth and the generation of electrostatic charges during rubbing process. Therefore, rubbing-free techniques for LC alignment are strongly required in LCD technology. In this experiment, PI was uniformly coated on indium-tin-oxide electrode substrates to form LC alignment layers using a spin-coating method and the PI layers were subsequently imidized at 433 K for 1 h. The thickness of the PI layer was set at 50 nm. The LC alignment layer surfaces were exposed to an $Ar^+$ ion-beam under various ion-beam energies. The antiparallel cells and twisted-nematic (TN) cells for the measurement of pretile angle and electro-optical characteristics were fabricated with the cell gap of 60 and $5\;{\mu}m$, respectively. The LC cells were filled with nematic LC (NLC, MJ001929, Merck) and were assembled. The NLC alignment capability on ion-beam-treated PI was observed using photomicroscope and the pretilt angle of the NLC was measured by the crystal-rotation method at room temperature. Voltage-transmittance (V-T) and response time characteristics of the ion-beam irradiated TN cell were measured by a LCD evaluation system.

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낙동강 상류 수역에서 남조류 발생과 천이패턴 - Aphanizomenon 속을 중심으로 - (Occurrence and Succession Pattern of Cyanobacteria in the Upper Region of the Nakdong River : Factors Influencing Aphanizomenon Bloom)

  • 류희성;박혜경;이혜진;신라영;천세억
    • 한국물환경학회지
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    • 제32권1호
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    • pp.52-59
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    • 2016
  • This study investigated the occurrences and succession patterns of harmful cyanobacteria, as well as environmental factors, during a 3-year period (September 2012 to August 2015) in the upper region of the Nakdong River around Sangju weir. A total of 27 cyanobacterial taxa were observed in this study, and classified into 26 species and 1 variety belonging to 11 genera, 5 families, and 3 orders. Cell density ranged from 24 to 42,001 cells/ml, with a geometric mean of 33 cells/ml, during the survey period. The dominant orders differed depending on the survey year; order Oscillatoriales in 2013, Chroococcales in 2014 and Nostocales in 2015. An Aphanizomenon bloom occurred in June 2015 at which time the highest cell density of 36,873 cells/ml was detected in the upper region of the Nakdong River, where as the Aphanizomenon spp. cell density (190-1,704 cells/ml) had been low prior to that time. An Aphanizomenon bloom also occurred at around the same time downstream in the Young River, a major inflow branch of the Nakdong River. The Aphanizomenon cell density along the Nakdong River increased markedly after joining of the YoungRiver, indicating that the Aphanizomenon bloom in the YoungRiver caused a bloom in the Nakdong River. Meteorological and environmental parameters, such as very low precipitation, higher water temperature, pH, and TP concentration, and lower TN/TP ratio, in May and June of 2015 than in 2013 and 2014 exerted marked effects on the Aphanizomenon bloom in June 2015 in the Young River.

스핀 코팅 공정에 따른 액정디스플레이용 폴리이미드 배향막 특성 분석 (Effect of Spin Coating Speed on Characteristics of Polyimide Alignment Layer for Liquid Crystal Display)

  • 김진아;최세훈;박홍규
    • 한국전기전자재료학회논문지
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    • 제35권1호
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    • pp.58-65
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    • 2022
  • The field of liquid crystal display (LCD) is constantly in the spotlight and the process of depositing an alignment layer in the LCD manufacturing process is very important to obtain excellent performance such as low-power driving and high-speed response to improve LCD performance. Therefore, research on liquid crystal (LC) alignment is being actively conducted. When manufacturing LCD, it is necessary to consider the effect of the alignment layer thickness as one of the factors affecting various LCD performances. In addition, previous studies confirmed the LC alignment characteristics correlate with the rotation speed in the spin coating process. Therefore, the electro-optical properties of the LCD were investigated by manufacturing a polyimide alignment layer by varying the rotation speed in the spin coating process in this study. It was confirmed that the thickness of the polyimide alignment layer was controlled according to the spin coating conditions. The average transmittances of anti-parallel LC cells at the spin coating speed of 2,500 rpm and 3,000 rpm are about 60%, which indicates that the LC cell has relatively higher performance. At the spin coating speed of 3,000 rpm, the voltage-transmittance curve of twisted nematic (TN) LC cell was below 1.5 V, which means that the TN LC cell operated at a low power. In addition, high-speed operating characteristics were confirmed with a response time of less than 30 ms. From these derived data, we confirmed that the ideal spin coating speed is 3,000 rpm. And these results provide an optimized polyimide alignment layer process when considering enhanced future LCD manufacturing.