• Title/Summary/Keyword: proteomic analysis

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Generation of a Mammalian Gene Expression Vector Using Bovine Viral Diarrhea Virus (Bovine Vira1 Diarrhea Virus를 이용한 포유동물세포 발현벡터의 개발)

  • 이영민
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.86-95
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    • 2002
  • As a result of genome projects, the research to elucidate the function of a protein of interest has recently been well-recognized. In order to facilitate functional genomics, a useful mammalian gene expression vector is required. Using an infectious CDNA clone of BVDV pNADLclns-, we have developed a mammalian gene expression vector. In this study, a replication-competent full-length infectious CDNA clone containing puremycin acetyltransferase (pac) gene (pNADLclns-/pac) was successfully generated. The viral RNA replication and viral protein NS3 synthesis were examined by detecting metabollically $^{32}P$-labelled genomic viral RNA and immunoblotting with a mouse anti-NS3 antibody. To generate viral replicon as an expression vector, we examine if the viral structural genes (C, E0, El, E2) are required for viral replication by deletion analysis. As a result, all of the structural proteins are dispensable for viral replication per se, but essential for infectious viral particle formation. Based on our deletion analysis, we have generated a replication-competent BVDV viral replicon (pNADLclns-/pac/${\Delta}S$), whose structural genes are all deleted. In addition to NADLclns- /pac/${\Delta}S$, NADLclns-/ luc/${\Delta}S$ viral replicon containing luciferase gene as a reporter was constructed and fecund to be replication-compotent in HeLa and BHK cells as well as MDBK cells. Therefore, BVDV viral replicon developed in our study will be a useful tool to express a protein of interest in various mammalian cells.

Proteomic Analysis and Growth Responses of Rice with Different Levels of Titanium Dioxide and UV-B (이산화티탄과 UV-B 수준에 따른 벼 생육과 프로테옴 해석)

  • Hong, Seung-Chang;Shin, Pyung-Gyun;Chang, An-Cheol;Lee, Ki-Sang;Lee, Chul-Won;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.1
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    • pp.69-80
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    • 2007
  • Among the photoactive semiconductors such as $TiO_2,\;ZnO,\;Fe_2O_3,\;WO_3,\;and\;CdSe,\;TiO_2$ is the most widely used as photocatalyst in different media, because of its lack of toxicity and stability. In this study, the effects of titanium dioxide were investigated to obtain the information of physiological change in rice plant. Light-adapted Chlorophyll flourescence index decreased and relative electron transport rate of rice leaves was activated by titanium dioxide under $2,400\;{\mu}mol\;m^{-2}\;s^{-1}$ PAR (Photosynthetic active radiation). Relative electron transport rate of rice leaf treated with titanium dioxide 10 ppm was high in order of $2,400\;{\mu}mol\;m^{-2}\;s^{-1}\;PAR,\;2,200\;{\mu}mol\;m^{-2}\;s^{-1}\;PAR,\;450\;{\mu}mol\;m^{-2}\;s^{-1}\;PAR$ and titanium dioxide 10 ppm (45.1%), control (32.4%), diuron 10 ppm (15.3%) under $2,400\;{\mu}mol\;m^{-2}\;s^{-1}\;PAR$. Titanium dioxide increased photosynthesis of the rice leaf under $13.6\;KJ\;m^{-2}\;day^{-1}$ UV-B only. With titanium dioxide 20 ppm, reduced UV-B ($0.15\;KJ\;m^{-2}\;day^{-1}$) intensity changed the induction of proteins and twenty-five proteins were identified. Among them, seventy proteins were up-regulated, four proteins were down-regulated and four proteins were newly synthesized. Function of these proteins was related to photosynthesis (52%), carbohydrate metabolism (4%), stress/defense (8%), secondary metabolism (4%), energy/electron transport (4%), and miscellaneous (28%).

Mitigation Effects of Foliar-Applied Hydrogen Peroxide on Drought Stress in Sorghum bicolor (과산화수소 엽면 처리에 의한 수수에서 한발 스트레스 완화 효과)

  • Shim, Doo-Do;Lee, Seung-Ha;Chung, Jong-Il;Kim, Min Chul;Chung, Jung-Sung;Lee, Yeong-Hun;Jeon, Seung-Ho;Song, Gi-Eun;Shim, Sang-In
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.2
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    • pp.113-123
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    • 2020
  • Global climatic change and increasing climatic instability threaten crop productivity. Due to climatic change, drought stress is occurring more frequently in crop fields. In this study, we investigated the effect of treatment with hydrogen peroxide (H2O2) before leaf development on the growth and yield of sorghum for minimizing the damage of crops to drought. To assess the effect of H2O2 on the growth of sorghum plant, 10 mM H2O2 was used to treat sorghum leaves at the 3-leaf stage during growth in field conditions. Plant height, stem diameter, leaf length, and leaf width were increased by 7.6%, 9.6%, 8.3% and 11.5%, respectively. SPAD value, chlorophyll fluorescence (Fv/Fm), photosynthetic rate, stomatal conductance, and transpiration rate were increased by 3.0%, 4.9%, 26.0%, 23.4% and 12.7%, respectively. The amount of H2O2 in the leaf tissue of sorghum plant treated with 10 mM H2O2 was 0.7% of the applied amount after 1 hour. The level increased to approximately 1.0% after 6 hours. The highest antioxidant activity measured by the Oxygen Radical Absorbance Capacity assay was 847.3 µmol·g-1 at 6 hour after treatment. However, in the well-watered condition, the concentration of H2O2 in the plant treated by the foliar application of H2O2 was 227.8 µmol·g-1 higher than that of the untreated control. H2O2 treatment improved all the yield components and yield-related factors. Panicle length, plant dry weight, panicle weight, seed weight per plant, seed weight per unit area, and thousand seed weight were increased by 8.8%, 18.0%, 24.4%, 24.7%, 29.9% and 7.1%, respectively. Proteomic analysis showed that H2O2 treatment in sorghum increased the tolerance to drought stress and maintained growth and yield by ameliorating oxidative stress.