• Title/Summary/Keyword: hre oxygen

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Analysis of Putative Downstream Genes of Arabidopsis AtERF71/HRE2 Transcription Factor using a Microarray (마이크로어레이를 이용한 애기장대 AtERF71/HRE2 전사인자의 하위 유전자 분석)

  • Seok, Hye-Yeon;Lee, Sun-Young;Woo, Dong-Hyuk;Park, Hee-Yeon;Moon, Yong-Hwan
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1359-1370
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    • 2012
  • Arabidopsis AtERF71/HRE2, a transcription activator, is located in the nucleus and is involved in the signal transduction of low oxygen and osmotic stresses. In this study, microarray analysis using AtERF71/HRE2-overexpressing transgenic plants was performed to identify genes downstream of AtERF71/HRE2. A total of 161 different genes as well as AtERF71/HRE2 showed more than a twofold higher expression in AtERF71/HRE2-overexpressing transgenic plants compared with wild-type plants. Among the 161 genes, 24 genes were transcriptional regulators, such as transcription factors and DNA-binding proteins, based on gene ontology annotations, suggesting that AtERF71/HRE2 is an upstream transcription factor that regulates the activities of various downstream genes via these transcription regulators. RT-PCR analysis of 15 genes selected out of the 161 genes showed higher expression in AtERF71/HRE2-overexpressing transgenic plants, validating the microarray data. On the basis of Genevestigator database analysis, 51 genes among the 161 genes were highly expressed under low oxygen and/or osmotic stresses. RT-PCR analysis showed that the expression levels of three genes among the selected 15 genes increased under low oxygen stress and another three genes increased under high salt stress, suggesting that these genes might be downstream genes of AtERF71/HRE2 in low oxygen or high salt stress signal transduction. Microarray analysis results indicated that AtERF71/HRE2 might also be involved in the responses to other abiotic stresses and also in the regulation of plant developmental processes.

Production of tissue-type plasminogen activator from immobilized CHO cells introduced hypoxia response element

  • Bae, Geun-Won;Kim, Hong-Jin;Kim, Gi-Tae;Kim, Ik-Yeong
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.257-260
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    • 2002
  • Dissolved oxygen level of cell culture media has a critical effect on cellular metabolism, which governs specific productivity of recombinant proteins and mammalian cell growth However, in the cores of cell aggregates or cell-immobilized beads, oxygen level frequently goes below a critical level. Mammalian cells have a number of genes induced in the lower level of oxygen, and the genes contain a common cis-acting element (-RCGTG-), hypoxia response element (HRE). By binding of hypoxia inducible factor-l (HIF-I) to the HRE, promoters of hypoxia inducible genes are activated, which is a survival mechanism. In this work, to develop a CHO cell capable of producing recombinant proteins in immobilization and high density cell culture efficiently, mammalian expression vectors containing human tissue-type plasminogen activator (t-PA) gene controlled by HRE were constructed and stably transfected into the CHO cells. In $Ba^{2+}$ -alginate immobilization culture, CHO/pCl/dhfr/2HRE-t-PA cells produced 2 folds higher recombinant t-PA activity than CHO/pCl/dhfrlt-PA cells without $CoCl_2$ treatment. Furthermore, in repeated fed batch culture, productivity of t-PA in immobilized CHO/pCI/dhfr/2HRE-t-PA cells was 121 ng/ml/day, total production of 0.968 mg/day at 11 days culture while CHO/pCIIdhfrlt-PA cells was 22.8 ng/ml/day. All these results indicate that HRE is very useful for the enhancement of protein productivity in mammalian cell cultures.

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High Productivity of t-PA in CHO Cells Using Hypoxia Response Element

  • Bae Gun-Won;Jeong Dae-Won;Kim Hong-Jin;Lee Gyun-Min;Park Hong-Woo;Choe Tae-Boo;Kang Seong-Man;Kim Ick-Young;Kim Ik-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.16 no.5
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    • pp.695-703
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    • 2006
  • The dissolved oxygen level of any cell culture environment has a critical effect on cellular metabolism. Specifically, hypoxia condition decreases cell viability and recombinant protein productivity. In this work, to develop CHO cells producing recombinant protein with high productivity, mammalian expression vectors containing a human tissue-type plasminogen activator (t-PA) gene with hypoxia response element (HRE) were constructed and stably transfected into CHO cells. CHO/2HRE-t-PA cells produced 2-folds higher recombinant t-PA production than CHO/t-PA cells in a $Ba^{2+}-alginate$ immobilized culture, and 16.8-folds in a repeated batch culture. In a non-aerated batch culture of suspension-adapted cells, t-PA productivity of CHO/2HRE/t-PA cells was 4.2-folds higher than that of CHO/t-PA cells. Our results indicate that HRE is a useful tool for the enhancement of protein productivity in mammalian cell cultures.

Spatio-temporal Distribution of Macrozoobenthos in the Three Estuaries of South Korea (우리나라 3개 하구역 대형저서동물 군집 시공간 분포)

  • LIM, HYUN-SIG;LEE, JIN-YOUNG;LEE, JUNG-HO;SHIN, HYUN-CHUL;RYU, JONGSEONG
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.24 no.1
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    • pp.106-127
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    • 2019
  • This study aims to understand spatio-temporal variations of macrozoobenthos community in Han River (HRE), Geum River (GRE), and Nakdong River estuaries (NRE) of Korea, sampled by National Survey of Marine Ecosystem. The survey was seasonally performed at a total of 20 stations for three years (2015-2017). Sediment samples were taken three times with van Veen grab of $0.1m^2$) areal size and sieved through a 1 mm pore size mesh on site. A total of 1,008 species were identified with 602 species in HRE, 612 in GRE, and 619 in NRE, showing similar number of species between estuaries. Mean density was $1,357ind./m^2$, showing the high in NRE ($1,357ind./m^2$), mid in GRE ($1,357ind./m^2$), and low in HRE ($1,127ind./m^2$). Mean biomass was $116.8g/m^2$, showing similar variations to density ($174.2g/m^2$ in NRE, $129.0g/m^2$ in GRE, $49.0g/m^2$ in HRE). Polychaeta dominated in number of species and density in three estuaries. Biomass-dominated taxon was Mollusca in HRE and GRE, and Echinodermata in NRE. Polychaetous species dominated all three estuaries over 4% of density, such as Dispio oculata, Heteromastus filiformis and Aonides oxycephala in HRE, Heteromastus filiformis and Scoletoma longifolia in GRE, and Pseudopolydora sp. and Aphelochaeta sp. in NRE, showing various density between estuaries. Community structure was determined by various environmental variables among estuaries such as mean grain size and sorting (HRE), salinity and mean grain size (GRE), and salinity, dissolved oxygen, loss on ignition and mud content (NRE). Our study demonstrates the application of different measures to manage ecosystems in three estuaries. HRE needs to alleviate sedimentary stressors such as sand mining, land-filling, dike construction. Management of GRE should be focused on fresh water control and sedimentary stressors. In NRE, monitoring of dominant benthos and process study on hypoxia occurrence in inner Masan Bay are necessary.

Generation of ROS by IgE-Dependent Histamine-Releasing Factor in RBL-2H3 Cells (RBL-2H3에서 IgE-dependent Histamine-releasing Factor에 의한 활성산소종 생성에 관한 연구)

  • Choo, Yee-Shin;Lee, Kyung-Iim
    • Microbiology and Biotechnology Letters
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    • v.33 no.3
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    • pp.231-235
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    • 2005
  • Histamine-releasing factors (HRFs) are soluble mediators that can release histamine and other mediators from basophils and mast cells and their activity can vary, depending on the type of IgE. The activity of HRFs is affected by the presence of IgE, although HRF is thought to bind to a specific receptor other than IgE. Until now, HRF signaling pathway including its receptor remains unclear in spite of numerous studies. Since there had been many reports about reactive oxygen species (ROS) as a signaling molecule rather than as a by-product of metabolism, we investigated the possibility of ROS as an intracellular messenger involved in HRF-mediated histamine degranulation. In RBL-2H3 cells, ROS was generated by HRF using $H_2O_2$-sensitive fluorescence of fluorescent 2', 7'-dichlorofluorescein ($H_2DCFDA$). These effects were blocked by anti-oxidant N-acetylcysteine (NAC). These results suggest that ROS generation could play a role as an intracellular messenger in histamine release by HRF.

Oxidation Behavior of Si3N4 by the Nitrided Pressureless Sintering (Nitrided Pressureless Sintering에 의해 제조된 Si3N4의 산화거동)

  • Han, In-Sub;Cheon, Sung-Ho;Jung, Yong-Hee;Seo, Doo-Won;Lee, Shi-Woo;Hong, Kee-Soeg;Woo, Sang-Kuk
    • Journal of the Korean Ceramic Society
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    • v.42 no.1
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    • pp.62-68
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    • 2005
  • Oxidtion behavior of $Si_{3}N_{4}$ ceramics with the different porosity by the Nitrided Pressureless Sintering (NPS) were investigated in pure oxygen gas atmosphere at 1000 to $1300^{circ}C$. The thickness of formed oxide film on the surface of silicon nitride ceramics was increased with oxidation time and temperature. The oxide film thickness of 5A5Y5Si and 5A5Y10Si specimens for 100 h at 1300^{circ}C$ was about 10 $\mu$m and 20 $\mu$m, respectively. The oxidation of 5A5Y5Si and 5A5Y10Si specimens follows the parabolic behavior with an apparent activation energy of 215 kJ/mol and 104 kJ/mol, respectively. The flexural strength of 5A5Y5Si specimens after oxidation test for 500 h at 1300^{circ}C were maintained as-received value of 500 ma. On the other hand, that of 5A5Y10Si specimens were decreased about 100 MPa in as-received value.