• Title/Summary/Keyword: Functional characterization

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Characterization of a Novel DWD Protein that Participates in Heat Stress Response in Arabidopsis

  • Kim, Soon-Hee;Lee, Joon-Hyun;Seo, Kyoung-In;Ryu, Boyeong;Sung, Yongju;Chung, Taijoon;Deng, Xing Wang;Lee, Jae-Hoon
    • Molecules and Cells
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    • v.37 no.11
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    • pp.833-840
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    • 2014
  • Cullin4-RING ubiquitin ligase (CRL4) is a family of multi-subunit E3 ligases. To investigate the possible involvement of CRL4 in heat stress response, we screened T-DNA insertion mutants of putative CRL4 substrate receptors that exhibited altered patterns in response to heat stress. One of the mutants exhibited heat stress tolerance and was named heat stress tolerant DWD1 (htd1). Introduction of HTD1 gene into htd1-1 led to recovery of heat sensitivity to the wild type level, confirming that the decrease of HTD1 transcripts resulted in heat tolerance. Therefore, HTD1 plays a negative role in thermotolerance in Arabidopsis. Additionally, HTD1 directly interacted with DDB1a in yeast two-hybrid assays and associated with DDB1b in vivo, supporting that it could be a part of a CRL4 complex. Various heat-inducible genes such as HSP14.7, HSP21, At2g03020 and WRKY28 were hyper-induced in htd1-1, indicating that HTD1 could function as a negative regulator for the expression of such genes and that these genes might contribute to thermotolerance of htd1-1, at least in part. HTD1 was associated with HSP90-1, a crucial regulator of thermotolerance, in vivo, even though the decrease of HTD1 did not affect the accumulation pattern of HSP90-1 in Arabidopsis. These findings indicate that a negative role of HTD1 in thermotolerance might be achieved through its association with HSP90-1, possibly by disturbing the action of HSP90-1, not by the degradation of HSP90-1. This study will serve as an important step toward understanding of the functional connection between CRL4-mediated processes and plant heat stress signaling.

Characterization of Insulation Finish Material Using Inorganic Wet Treatment Fly Ash (무기성 습식 처리 플라이애시를 활용한 단열 외피 마감재의 특성 평가)

  • Ryu, Hwa-Sung;Shin, Sang-Heon;Song, Sung-Young;Kim, Deuak-Mo
    • Journal of the Korean Recycled Construction Resources Institute
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    • v.7 no.4
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    • pp.389-394
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    • 2019
  • In this study, a functional inorganic insulation as TiO2 and inorganic wet fly ash were used to evaluate the physical performance and thermal environment of an exterior finish that can improve the thermal environment of a building. The performance evaluation of the finish was based on the KS F 4715 thin coating material and the thermal environment. When TiO2 was added, the physical performance was lowered at 10% or more, and the inorganic wet-treated fly ash increased the physical performance by 10%. In the thermal environmental evaluation, the surface temperature reduction effect of the inorganic wet-treated fly ash was low, but when used in combination with TiO2, it was effective to reduce the surface temperature and the internal temperature. As a result, the optimum combination of TiO2 and inorganic wet-treated fly ash for thermal environment control was found to be optimal when 5% of each mixture was used.

Construction and Characterization of a cDNA Library from the Camelina sativa L. as an Alternative Oil-Seed Crop (신 바이오디젤 원료 작물인 Camelina의 cDNA library 제작 및 유전자 특성)

  • Park, Won;Jang, Young-Seok;Ahn, Sung-Ju
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.55 no.2
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    • pp.151-158
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    • 2010
  • Camelina sativa L., known as popular names "gold-of-pleasure" or "false flax" is an alternative oilseed crop that can be grown under different climatic and soil conditions. Up to date, however, the genomic information of Camelina has not been studied in detail. Therefore, a cDNA library was constructed and characterized from young leaves. The constructed cDNA library incorporated of 1334 cDNA clones and the size of the insertion fragments average was 736 base pair. We generated a total of 1269 high-quality expressed sequence tags (ESTs) sequences. The result of cluster analysis of EST sequences showed that the number of unigene was 851. According to subsequent analysis, the 476 (55.9%) unigenes were highly homologous to known function genes and the other 375 (44.1%) unigenes were unknown. Remaining 63 (7.4%) unigenes had no homology with any other peptide in NCBI database, indicating that these seemed to be novel genes expressed in leaves of Camelina. The database-matched ESTs were further classified into 17 categories according to their functional annotation. The most abundant of categories were "protein with binding function or cofactor requirement (27%)", "metabolism (11%)", "subcellular localization (11%)", "cellular transport, transport facilities and transport routes (7%)", "energy (6%)", "regulation of metabolism and protein function (6%)". Our result in this study provides an overview of mRNA expression profile and a basal genetic information of Camelina as an oilseed crop.

Spectroscopic Characterization and Seasonal Distribution of Aquatic Humic Substances Isolated from Han River Water (한강원수로부터 분리된 수중휴믹물질의 계절적 분포와 분광학적 특성분석)

  • Kim, Hyun-Chul;Lee, Seock-Heon;Kim, Kyung-Ju;Yu, Myong-Jin
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.5
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    • pp.540-547
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    • 2007
  • Humic substances(HS) from Han River water was physic-chemically isolated by fractionational methods to investigate the seasonal distribution and to characterize the properties compared with intrinsic humic materials. Various HS samples were analyzed by element, Fourier transform infrared(FT-IR), proton nuclear magnetic resonance$(^1H-NMR)$ and fluorescence analyzers. The portion of HS from Han River water(HRHS) was 47.0% on the average, however it appeared that rainfall event brought about higher fraction of HS in Han River water by the periodic investigation. Aromaticity and humification degree of the HRHS were relatively lower than those of intrinsic humic materials originated from decomposing vegetation. FT-IR, $^1H-NMR$ and fluorescence spectroscopy showed the distinct differences between HRHS and intrinsic humic materials. Commercial humic materials could not represent structural and functional characteristics of local HS. The fluorescence spectroscopy, a relatively simple measurement, was found most useful tool to estimate humification degree for humic materials from particular sources.

Food Component Characteristics of Tang from Conger Eel By-products (붕장어 부산물로 제조한 붕장어탕의 식품학적 특성)

  • Heu, Min-Soo;Lee, Take-Sang;Kim, Hye-Suk;Jee, Seung-Joon;Lee, Jae-Hyoung;Kim, Hyung-Jun;Yoon, Min-Seok;Park, Shin-Ho;Kim, Jin-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.4
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    • pp.477-484
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    • 2008
  • For the effective use of the conger eel by-products, such as head and frame, Tang, which is the Korean-type soup, from conger eel by-products (TCEB) was developed and its food component characterization was compared with that of commercial Chueotang, loach Tang. According to the results of viable cells and coliform group of TCEB heated at $115^{\circ}C$ for various times, the reasonable $F_0$ value was 8 min. The proximate composition of TCEB was 90.7% for the moisture, 4.8% for the protein, 2.6% for the lipid, and 1.5% for the ash. The extractive-nitrogen content of TCEB was 243.1 mg/100 g, which was higher than that of commercial Chueotang, 208.0 mg/100 g. The total amino acid content of TCEB was 4,310 mg/100 g and its major amino acids were glutamic acid (637.3 mg/100 g, 14.8%), glycine (409.1 mg/100 g, 9.5%) and alanine (404.4 mg/100 g, 9.3%). TCEB was not felt in the sensual fish odor and its sensual taste was good. The health functional properties for health of TCEB were 1.29 as a PF (protection factor) for antioxidative activity and 39.4% for angiotensin Ⅰ converting enzyme (ACE) inhibiting activity.

Synthesis and Characterization of Poly(ethylene glycol) Grafted Polysuccinimide (폴리(에틸렌 글리콜)이 결합된 Polysuccinimide의 합성과 특성)

  • Lim, Nak-Hyun;Lee, Ha-Young;Kim, Moon-Suk;Khang, Gil-Son;Lee, Hai-Bang;Cho, Sun-Hang
    • Polymer(Korea)
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    • v.29 no.1
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    • pp.36-40
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    • 2005
  • Poly(amino acid) derivatives have been widely investigated as a drug carrier in drug delivery system. Particularly,polysuccinimide (PSI) is one of the most promising drug carriers since it possesses suitable physicochemical characteristics for development of macromolecular prodrugs, due to biocompatibility and biodegradability. In this study, we deal with the synthesis of polyaspartamide having various functional groups such as methoxy-poly(ethylene glycol) (MPEG) via ring closing of PSI. PSI was synthesized by polyonensation polymerization of spartic acid. The variety of average molecular weight was confirmed with reacion time and catalyst content to observe the optimum condition of synthesis. MPEG, hydrophilic chain, was bonded to fabricate polymeric micell composed of hydrophilic and hydrophobic polymer. All materials were characterized by 1H-NMR, FT-IR and GPC. In addition, the formation of nanoparticle micelle as drug carrier were also examined. Micelle size was measured by ELS and AFM. The functionalized polysparamide formed nanoparticle micelle whose size ranged from 90 to 130 nm. In conclusion, we prepared polyaspartamide functionalized with PEG examined the possibility as drug carriers.

Identification and Characterization of the Aquaporin Gene aqpA in a Filamentous Fungus Aspergillus nidulans (사상성 진균 Aspergillus nidulans에서 아쿠아포린 유전자 aqpA의 분리 및 분석)

  • Oh, Dong-Soon;Lu, Han-Yan;Han, Kap-Hoon
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.295-301
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    • 2011
  • Aquaporin is a water channel protein, which is classified as Major Intrinsic Protein (MIP), found in almost all organisms from bacteria to human. To date, more than 200 members of this family were identified. There are two major categories of MIP channels, orthodox aquaporins and aquaglyceroporins, which facilitate the diffusion across biological membranes of water or glycerol and other uncharged compounds, respectively. The full genome sequencing of various fungal species revealed 3 to 5 aquaporins in their genome. Although some functions of aquaporins found in yeast were characterized, however, no functional characteristics were studied so far in filamentous fungi, including Aspergillus sp. In this study, one orthodox aquaporin homolog gene, aqpA, and four aquaglyceroporin homologs, aqpB-E, in a model filamentous fungus Aspergillus nidulans were identified and the function of the aqpA gene was characterized. Knock-out of the aqpA gene didn't show any obvious phenotypic change under the osmotic stress, indicating that the function of the gene does not involved in the osmotic stress response or the function could be redundant. However, the mutant showed antifungal susceptibility resistance phenotype, suggesting that the function of the aqpA gene could be involved in sensing the antifungal substances rather than the osmotic stress response.

Bacillus subtilis를 이용한 대두 발효식품의 혈전용해능

  • Jeong, Yeong-Gi
    • Proceedings of the Korean Society of Life Science Conference
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    • 2001.06a
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    • pp.67-86
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    • 2001
  • A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.

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Identification and Characterization of Alternative Promoters of the Rice MAP Kinase Gene OsBWMK1

  • Koo, Sung Cheol;Choi, Man Soo;Chun, Hyun Jin;Park, Hyeong Cheol;Kang, Chang Ho;Shim, Sang In;Chung, Jong Il;Cheong, Yong Hwa;Lee, Sang Yeol;Yun, Dae-Jin;Chung, Woo Sik;Cho, Moo Je;Kim, Min Chul
    • Molecules and Cells
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    • v.27 no.4
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    • pp.467-473
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    • 2009
  • Our previous study suggested that OsBWMK1, a gene which encodes a member of the rice MAP kinase family, generates transcript variants which show distinct expression patterns in response to environmental stresses. The transcript variants are generated by alternative splicing and by use of alternative promoters. To test whether the two alternative promoters, pOsBWMK1L (promoter for the OsBWMK1L splice variant) and pOsBWMK1S (promoter for the OsBWMK1S splice variant), are biologically functional, we analyzed transgenic plants expressing GUS fusion constructs for each promoter. Both pOsBWMK1L and pOsBWMK1S are biologically active, although the activity of pOsBWMK1S is lower than that of pOsBWMK1L. Histochemical analysis revealed that pOsBWMK1L is constitutively active in most tissues at various developmental stages in rice and Arabidopsis, whereas pOsBWMK1S activity is spatially and temporally restricted. Furthermore, the expression of pOsBWMK1S::GUS was upregulated in response to hydrogen peroxide, a plant defense signaling molecule, in both plant species. These results suggest that the differential expression of OsBWMK1 splice variants is the result of alternative promoter usage and, moreover, that the mechanisms controlling OsBWMK1 gene expression are conserved in both monocot and dicot plants.

Preparation and Characterization of Cu/MCM-41 Mesoporous Catalysts for NO Removal (Cu/MCM-41 메조포러스 촉매 제조 및 NO 제거 특성)

  • Park, Soo-Jin;Cho, Mi-Hwa;Kim, Seok;Kwon, Soo-Han
    • Applied Chemistry for Engineering
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    • v.16 no.6
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    • pp.737-741
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    • 2005
  • In this study, the effect of copper content on the NO removal efficiency by Cu/MCM-41 has been investigated. MCM-41 was prepared by hydrothermal synthesis using a gel mixture of colloidal silica solution and cetyltrimethylammonium. Cu/MCM-41 was manufactured with copper content (5, 10, 20, and 40%) in Cu(II) acetylacetonate. The surface properties of MCM-41 were investigated by using pH, XRD, and FT-IR analyses. $N_2/77K$ adsorption isotherm characteristics, including the specific surface area and micropore volume were studied by BET's equation and Boer's t-plot methods. NO removal efficiency was confirmed by gas chromatography technique. From the experimental results, the MCM-41 was analyzed to have the surface functional groups of Si-OH and Si-O-Si and the characteristic diffraction lines (100), (110), (200), and (210) corresponding to a hexagonal arrangement structure. The copper content supported on MCM-41 appeared to increase the NO removal efficiency in spite of decreasing the specific surface areas or micropore volumes. Consequently, it was found that the copper content in Cu/MCM-41 played an important role in improving the NO removal efficiency, which was mainly attributed to the catalytic reactions.