• 제목/요약/키워드: stable cells

검색결과 967건 처리시간 0.027초

Efficient transgene expression system using a cumate-inducible promoter and Cre-loxP recombination in avian cells

  • Park, Tae Sub;Kim, Si Won;Lee, Jeong Hyo
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권6호
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    • pp.886-892
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    • 2017
  • Objective: Transgenic technology is widely used for industrial applications and basic research. Systems that allow for genetic modification play a crucial role in biotechnology for a number of purposes, including the functional analysis of specific genes and the production of exogenous proteins. In this study, we examined and verified the cumate-inducible transgene expression system in chicken DF1 and quail QM7 cells, as well as loxP element-mediated transgene recombination using Cre recombinase in DF1 cells. Methods: After stable transfer of the transgene with piggyBac transposon and transposase, transgene expression was induced by an appropriate concentration of cumate. Additionally, we showed that the transgene can be replaced with additional transgenes by co-transfection with the Cre recombinase expression vector. Results: In the cumate-GFP DF1 and QM7 cells, green fluorescent protein (GFP) expression was repressed in the off state in the absence of cumate, and the GFP transgene expression was successfully induced in the presence of cumate. In the cumate-MyoD DF1 cells, MyoD transgene expression was induced by cumate, and the genes controlled by MyoD were upregulated according to the number of days in culture. Additionally, for the translocation experiments, a stable enhanced green fluorescent protein (eGFP)-expressing DF1 cell line transfected with the loxP66-eGFP-loxP71 vector was established, and DsRed-positive and eGFP-negative cells were observed after 14 days of co-transfection with the DsRed transgene and Cre recombinase indicating that the eGFP transgene was excised, and the DsRed transgene was replaced by Cre recombination. Conclusion: Transgene induction or replacement cassette systems in avian cells can be applied in functional genomics studies of specific genes and adapted further for efficient generation of transgenic poultry to modulate target gene expression.

가정용연료전지 전기적 안전성 평가 기법 개발 (Development of Electrical Safety Evaluation Method about PEMFC 1kW)

  • 한운기;박찬엄;정진수;고원식
    • 한국신재생에너지학회:학술대회논문집
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    • 한국신재생에너지학회 2011년도 춘계학술대회 초록집
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    • pp.99-99
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    • 2011
  • Fuel cell systems are a completely different form of electricity source that has been used so far and is an aggregation of multiple technologies with multidisciplinary features that can be operated safely only when gas and electrical safety are being considered. Since fuel cells generate through electrochemical reactions there are difficulties in ensuring electricity safety, power quality assessment, effective control and reliability standards for system faults using conventional inspection techniques and even though they are necessary as a primary means for reduction of CO2 owing to the Climate Convention, electrical safety assessment and measures are required for the prevention of faults in residential facilities. Although small-scaled distributed power supplies can be utilized as important means of peak control and energy management measures, research is required for observing the effects on the system and the development of inspection technology to ensure stable operation, and the electrical safety of residential fuel cell systems need to be assessed and the problems derived for establishing electrical safety standards. From the year 2002, Japan has established laws on technical safety standards and development and rules on the product specifications and standards for the industrialization of hydrogen fuel cells. Also, a lot of effort have been made for the commercialization of fuel cells by building one-stop certification services. Internationally, the IEC TC 105 has established international standards based on fuel cells. In order to protect the national interest, the country should be able to respond accordingly meet global standards. In fact, in Korea, to comply with the international trend, Korea Energy Management Corporation is establishing a certified agenda for fuel cells and Korean Agency for Technology and Standards is enacting technical standards for fuel cells. The current terms of fuel cells are that research has been focused more on the quality and performance of manufactured products rather than stable power operation and maintenance over time. In this paper, by considering the household fuel cell as a power device, the safety standards of the fuel cell system for a reliable operation with the existing power system is being proposed.

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최적의 luminescence 신호 분석을 위한 유전자 전달 방법의 비교연구 (Comparative studies of various transfection processes for the optimal luminescence signal analysis)

  • 박서현;이승호
    • 한국산학기술학회논문지
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    • 제17권11호
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    • pp.640-647
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    • 2016
  • 형광 간섭 현상을 최소화시켜 상대적으로 민감한 측정이 가능한 aequorin기반 luminescence기술은 $G_{{\alpha}16}$ 단백질 도입을 통해 세포 내부의 칼슘 이동 신호를 감지하여 G 단백질 결합 수용체(G protein-coupled receptor, GPCR)의 기능 분석을 가능하게 하는 세포 기반 분석 기술로 수용체 및 G 단백질 유전자 전달의 최적화 과정이 필수적이다. 본 연구를 위해 corticotropin releasing factor receptor subtype 2(CRF2) 수용체를 모델 시스템으로 CRF2와 $G_{{\alpha}16}$ 단백질이 구축된 세 가지 안정화 세포주를 제작하였고, 이들을 이용한 서로 다른 세 가지 조건의 임시 발현 세포주에서 작용제(sauvagine)와 길항제(K41498)의 반응성을 분석하여 최적의 유전자 전달 방법을 도출하고자 하였다. 그 결과 sauvagine 및 K41498의 농도에 따른 반응에서 CRF2-$G_{{\alpha}16}$ 안정화 세포주가 임시 발현 세포주보다 10배 이상의 유효신호 비율을 나타내었고(z'=0.77) 임시 발현 세포주의 경우 $G_{{\alpha}16}$의 안정화 발현 이후에 CRF2를 전달하는 경우가 다른 임시 발현 조건보다 2배 이상 높은 효율을 보였다(z'=0.84). 따라서 임시 유전자 전달 기술을 GPCR 세포 기능 분석 시스템에 활용할 경우 $G_{{\alpha}16}$ 단백질에 대한 안정화 세포주를 우선적으로 구축하고, 목표하는 다양한 수용체들을 단계적으로 발현시키는 것이 최선의 방법이라 판단된다.

Establishment of Mouse Pluripotent Stem Cells Generated from Primordial Germ Cells

  • Shim, Sang-Woo;Song, Sang-Jin;Hosup Shim;Lee, Bo-Yon;Huh, Choo-Yup;Hyuk Song;Chung, Kil-Saeng;Lee, Hoon-Taek
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2004년도 춘계학술발표대회
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    • pp.276-276
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    • 2004
  • Pluripotent stem cells have been generated from two embryonic sources. ES cells are generated from ICM of blastocyst stage embryos, and embryonic germ (EG) cells are generated from primordial germ cells (PGCs). Both ES and EG cells are pluripotent and present important characteristics such as high levels of alkaline phosphatase (AP) activity, multi-cellular colony formation, normal and stable karyotypes, continuously passaging ability, and the capability of differentiation into all three embryonic germ layers. (omitted)

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An Increased Proportion of Apoptosis in CD4+ T Lymphocytes Isolated from the Peripheral Blood in Patients with Stable Chronic Obstructive Pulmonary Disease

  • Ju, Jinyung
    • Tuberculosis and Respiratory Diseases
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    • 제81권2호
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    • pp.132-137
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    • 2018
  • Background: The pathophysiology of chronic obstructive pulmonary disease (COPD) includes inflammation, oxidative stress, an imbalance of proteases and antiproteases and apoptosis which has been focused on lately. Abnormal apoptotic events have been demonstrated in both epithelial and endothelial cells, as well as in inflammatory cells including neutrophils and lymphocytes in the lungs of COPD patients. An increased propensity of activated T lymphocytes to undergo apoptosis has been observed in the peripheral blood of COPD patients. Therefore, the apoptosis of T lymphocytes without activating them was investigated in this study. Methods: Twelve control subjects, 21 stable COPD patients and 15 exacerbated COPD patients were recruited in the study. The T lymphocytes were isolated from the peripheral blood using magnetically activated cell sorting. Apoptosis of the T lymphocytes was assessed with flow cytometry using Annexin V and 7-aminoactinomycin D. Apoptosis of T lymphocytes at 24 hours after the cell culture was measured so that the T lymphocyte apoptosis among the control and the COPD patients could be compared. Results: Stable COPD patients had increased rates of $CD4^+$ T lymphocyte apoptosis at 24 hours after the cell culture, more than the $CD4^+$ T lymphocyte apoptosis which appeared in the control group, while the COPD patients with acute exacerbation had an amplified response of $CD4^+$ T lymphocyte apoptosis as well as of $CD8^+$ T lymphocyte apoptosis at 24 hours after the cell culture. Conclusion: Stable COPD patients have more apoptosis of $CD4^+$ T lymphocytes, which can be associated with the pathophysiology of COPD in stable conditions.

안정적인 SOFC 운전을 위한 디젤 개질기 내 미반응 저탄화수소 제거법 (Methodology for removing unreacted low-hydrocarbons in diesel reformate for stable operation of solid oxide fuel cells)

  • 윤상호;배중면;이상호
    • 한국신재생에너지학회:학술대회논문집
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    • 한국신재생에너지학회 2009년도 춘계학술대회 논문집
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    • pp.773-776
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    • 2009
  • In this paper, new concept of the diesel fuel processing is introduced for the stable operation of solid oxide fuel cells (SOFCs). Heavier hydrocarbons than $CH_4$, such as ethylene, ethane, propane, and etc., induce the carbon deposition on anode of SOFCs. In the reformate of heavy hydrocarbons (diesel, gasoline, kerosene, and JP-8), concentration of ethylene is usually higher than low hydrocarbons such as ethane, propane, and butane. So, removal of low hydrocarbons (over C1-hydrocarbons), especially ethylene, at the reformate gases is important for stable operation of SOFCs. New methodology as named "post-reformer" is introduced for removing the low hydrocarbons at the reformate gas stream. Catalyst of the NECS-PR4 is selected for post-reforming catalyst because the catalyst of NECS-PR4 shows the high selectivity for removing low hydrocarbons and achieving the high reforming efficiency. The diesel reformer and post-reformer are continuously operated for about 200 hours as integrated mode. The reforming performance is not degraded and low hydrocarbons in the diesel reformate are completely removed.

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UV-조사 수정란 내로 이식한 유전자 변화 배반엽 세포의 재구성 (Recolonization of Transfected Blastodermal Cells in Developing Embryos after Transferring into UV-irradiated Fertilized Hen′s Egg)

  • 이기석;이황;김기동;박성수;이상호
    • 한국가금학회지
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    • 제27권2호
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    • pp.155-161
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    • 2000
  • Unfortunately, there is no technique which is stable and repetitive to produce transgenic chicken, although various ways of gene transfer including PGC-and embryonic cell-mediated gene transfer, DNA microinjection, virus inoculation and sperm cells have been employed. The aims of this study were 세 develop and establish such a stable, repetitive and efficient way of gene transfer giving a faithful gene expression during development after the reconstruction of embryo in an UV-irradiated egg. A dual reporter plasmid (pJJ9), a fusion gene containing lacZ and GFP driven by a CMV promoter was used to exploit either merits of both reporting markers. lacZ with strong signal or GFP with vital marking. Electroporated embryonic blastodermal cells (EBCs) in the presence of the pJJ9 DNA faithfully showed 377 bp PCR product and lacZ or GFP expressions in the identical cells in vitro of in vivo. Furthermore, analyses of expression pattern of the foreign DNA demonstrated that microinjected EBCs cells into the UV-irradiated recipient egg should participate in normal developmental process, for example, proliferation and differentiation into various tissues. Thirty percentages of the manipulated eggs showed lacZ expression in their tissues. These results together with the specific procedures used in this study should facilitate avian transgenesis.

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ESTABLISHMENT OF BIOASSAY TO DETECT ESTROGENIC FLAVONOIDS USING STABLE MCF-7-ERE CELL AND MCF-7 CELL PROLIFERASTION ASSAY

  • Joung, Ki-Eun;Kim, Yeo-Woon;Sheen, Yhun-Yhong
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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    • pp.113-113
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    • 2001
  • Stable MCF-7-ERE cells, in which pERE-Luc reporter gene has been stably integrated into the genome of the MCF-7 cells, were used to detect the estrogenic activity of various dietary flavonoids in either pure chemical or mixtures. Estradiol (E2) induced luciferase activity in dose dependent manner and this activity was inhibited by tamoxifen (Tam) concomitant treatment. A large series of flavonoids showed estrogenic activities, corresponding to EC5O values between 0.2 and 9 microM and their mixtures didn't show additive or synergistic effects. And we could find some structure and activity relationship. First, 4-methoxylation and catechol structure decreased estrogenic activities. Second, hydroxylation of 3 position reduced estrogenic effect. Third glycosides of flavonoids showed weak estrogenic activity or no activity. Interestingly, when tested at high concentrations, genistein, kaempferol, biochanin A and chrysin elicited luciferase induction higher than that of the maximum induction by estradiol. And these effects of genistein and kaempferol could not be fully inhibited with tamoxifen

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Improving the Stability of Series-Connected Solid Oxide Fuel Cells by Modifying the Electrolyte Composition

  • Kim, Young Je;Lim, Hyung-Tae
    • Journal of Electrochemical Science and Technology
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    • 제12권1호
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    • pp.159-165
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    • 2021
  • YSZ based anode supported solid oxide fuel cells (SOFCs) were prepared, and two cells with different electrolyte thicknesses were connected in series for the simulation of a cell-imbalanced fuel cell stack. Pure YSZ cells in a series connection exhibited a rapid degradation when a thick electrolyte cell was operated under a negative voltage. On the other hand, ceria added-YSZ cells in a series connection were stable under similar operating conditions, and the power density and impedance were about the same as those before tests. The improved stability was due to the reduction of internal partial pressure in the electrolyte by locally increasing the electronic conduction. Thus, we propose a new protection method, i.e., the local addition of ceria in the YSZ electrolyte, to extend the lifetime of a cell-imbalanced SOFC stack.