• Title/Summary/Keyword: SodC

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Development and Characterization of Expression Vectors for Corynebacterium glutamicum

  • Lee, Jinho
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.70-79
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    • 2014
  • In an attempt to develop a variety of expression vector systems for Corynebacterium glutamicum, six types of promoters, including $P_{tac}$, $P_{sod}$, $P_{sod}$ with a conserved Shine-Dalgarno (SD) sequence from C. glutamicum, $P_{ilvC}$, $P_{ilvC}$ with a conserved SD-1 ($P_{ilvC-M1}$), and $P_{ilvC}$ with a conserved SD-2 ($P_{ilvC-M2}$), were cloned into a modified shuttle vector, pCXM48. According to analysis of promoter strength by quantitative reverse transcription PCR, $P_{sod}$ and $P_{sod-M}$ were superior to tac and ilvC promoters in terms of transcription activity in C. glutamicum. All of the promoters have promoter activities in Escherichia coli, and $P_{sod-M}$ displayed the highest level of transcriptional activity. The protein expression in constructed vectors was evaluated by measuring the fluorescence of green fluorescent protein (GFP) and SDS-PAGE. C. glutamicum harboring plasmids showed GFP fluorescence with an order of activity of $P_{ilvC}$ > $P_{ilvC-M1}$ > $P_{sod}$ > $P_{ilvC-M2}$ > $P_{sod-M}$, whereas all plasmids except pCSP30 with $P_{sod}$ displayed fluorescence activities in E. coli. Of them, the strongest level of GFP was observed in E. coli with $P_{sod-M}$, and this seems to be due to the introduction of the conserved SD sequence in the translational initiation region. These results demonstrate that the expression vectors work well in both C. glutamicum and E. coli for the expression of target proteins. In addition, the vector systems harboring various promoters with different strengths, conserved SD sequences, and multiple cloning sites will provide a comfortable method for cloning and gene expression, and consequently contribute to the metabolic engineering of C. glutamicum.

Isolation of Superoxide Dismutase cDNAS from an Weedy Rice Variety and Transformation of a Cultivated Rice Variety (잡초성벼의 superoxide dismutase cDNA cloning과 재배벼로의 형질전환)

  • Park, Sang-Gyu;Park, Jong-Suk;Lee, Seung-In;Suh, Suk-Chul;Kim, Byung-Keuk;Jo, Youl-Lae;Suh, Hak-Soo
    • Korean Journal of Environmental Agriculture
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    • v.21 no.2
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    • pp.156-161
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    • 2002
  • Two different cDNA clones for superoxide dismutase (SOD) were isolated from an weedy rice variety (Oryza sativa, cv. Bhutan14Ad) and were introduced into a cultivated rice variety (Oryza sativa, cv. Nakdong) in order to develop the environmental stress-resistant rice plants. Sequence analysis of the cloned cDNAS indicated that the deduced amino acid sequence of SOD-A is 88.4% identical to that of SOD-B. Furthermore, the nucleotide sequence of SOD-A is 99.3% identical to that of a Cu/Zn SOD gene of Oryza sativa (GenBank accession No. L36320). The nueleotide sequence of SOD-B was identical to that of the previously published SOD gene (Accession No. D01000). A cultivated rice variety, Nakdong-byeo, was transformed with chimeric SOD genes containing a actin promoter of rice and pin2 terminator using a particle bombardment technique. Transformed calli were selected on an selection medium containing phosphinothricin (PPT). Transgenic rice plants were regenerated from the PPT-resistant calli. PCR analysis with genomic DNAs from transgenic plants revealed that transgenes are introduced into rice genome.

Identification and Characterization of SOD Isoenzymes in Acanthopanax koreanum Plants (섬오갈피나무에서 SOD Isoenzyme의 식별 및 특성규명)

  • 오순자;박영철;김응식;고석찬
    • Korean Journal of Plant Resources
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    • v.12 no.3
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    • pp.234-239
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    • 1999
  • The isoenzyme patterns and activities of superoxide dismutase(SOD) were investigated from leaves of Araliaceae plants. Of the eight isoenzymes, two isoenzymes(SOD 4 and SOD 6) were prevalent to leaves of Araliaceae plants. The patterns of these two isoenzymes were most various in the leaves of Acanthopanax senticosus for. inermis, while their activity was highest in the leaves of A. koreanum. These two isoenzymes were respectively identified as Fe-SOD and CuZn-SOD, based on selective inhibition with KCN or$H_2O_2$. The SOD isoenzyme patterns did not differed among stem barks, root barks and leaves of A. koreanum. However, the activities of Fe-SOD and CuZn-SOD were higher in the root bark and in leaves, respectively. Both of Fe-SOD and CuZn-SOD were stable for 1 hr at 30-4$0^{\circ}C$, while unstable above 5$0^{\circ}C$.

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Genomic Structure of the Cu/Zn Superoxide Dismutase(SOD1) Gene from the Entomopathogenic Fungus, Cordyceps pruinosa

  • Park, Nam Sook;Jin, Byung Rae;Lee, Sang Mong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.39 no.2
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    • pp.67-73
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    • 2019
  • The genomic structure of the Cu/Zn superoxide dismutase (SOD1) gene from the entomopathogenic fungus, Cordyceps pruinosa was characterized. The SOD1 gene of C. pruinosa spans 947 nucleotides and consisted of four exons encoding for 154 amino acids and three introns. Four exons of the SOD1 gene are composed of 13, 331, 97 and 20 nucleotides respectively. Homology search of amino acid sequences of the SOD1 gene of C. pruinosa with another 13 fungi species showed higher sequence similarity of 69% ~ 95% and had the most highest sequence identity of 95% with Beauveria bassiana and Cordyceps militaris, which can easely infect domesticated Bombyx mori and another wild lepidopteran species in artificial or natual manner of infection. This SOD1 gene sequence showed copper, zinc and beta-barrel fold sites. Homology search showed that the Cu/Zn SOD1 gene from the entomopathogenic fungus, C. pruinosa is an orthologous gene homolog present in different species of organism whose ancestor predates the split between the relating species. In addition, C. pruinosa SOD1 gene is placed together within the ascomycetes group of fungal clade. From these results it is concluded that C. pruinosa SOD1 gene is orthologous gene having the same or very similar functions with a common evolutionary ancestor.

Molecular Cloning and Expression of Sequence Variants of Manganese Superoxide Dismutase Genes from Wheat

  • Baek, Kwang-Hyun;Skinner, Daniel Z.
    • Korean Journal of Environmental Agriculture
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    • v.29 no.1
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    • pp.77-85
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    • 2010
  • Reactive oxygen species (ROS) are very harmful to living organisms due to the potential oxidation of membrane lipids, DNA, proteins, and carbohydrates. transformed E.coli strain QC 871, superoxide dismutase (SOD) double-mutant, with three sequence variant MnSOD1, MnSOD2, and MnSOD3 manganese superoxide dismutase (MnSOD) gene isolated from wheat. Although all QC 871 transformants grown at $37^{\circ}C$ expressed mRNA of MnSOD variants, only MnSOD2 transformant had functional SOD activity. MnSOD3 expressed active protein when grown at $22^{\circ}C$, however, MnSOD1 did not express functional protein at any growing and induction conditions. The sequence comparison of the wheat MnSOD variants revealed that the only amino acid difference between the sequence MnSOD2 and sequences MnSOD1 and 3 is phenylalanine/serine at position 58 amino acid. We made MnSOD2S58F gene, which was made by altering the phenylalaine to serine at position 58 in MnSOD2. The expressed MnSOD2S58F protein had functional SOD activity, even at higher levels than the original MnSOD2 at all observed temperatures. These data suggest that amino acid variation can result in highly active forms of MnSOD and the MnSOD2S58F gene can be an ideal target used for transforming crops to increase tolerance to environmental stresses.

Changes of Vitamin C and Superoxide Dismutase(SOD)-like Activity of Persimmon Leaf Tea by Processing Method and Extraction Condition (감잎의 처리방법과 추출조건에 따른 감잎차의 Vitamin C와 Superoxide Dismutase(SOD) 유사활성의 변화)

  • Park, Yun-Joo;Kang, Myung-Hee;Kim, Jong-Ik;Park, Ock-Jin;Lee, Mee-Sook;Jang, Hae-Dong
    • Korean Journal of Food Science and Technology
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    • v.27 no.3
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    • pp.281-285
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    • 1995
  • The persimmon leaf tea was produced from persimmon leaves by three different methods (conventional, steamed, fermented) and the changes of total vitamin C and superoxide dismutase(SOD)-like activity were investigated. The amount of total vitamin C was the highest in fermented persimmon leaf tea which was 47% of total vitamin C of raw persimmon leaves. The SOD-like activity of conventional and fermented persimmon leaf teas were a little higher than that of steamed. The total vitamin C of steamed persimmon leaf tea was decreased slowly as the extraction time was increased. That of fermented persimmon leaf tea was increased generally at $60^{\circ}C\;and\;70^{\circ}C$, and increased until 5min and then decreased at $80^{\circ}C\;and\;90^{\circ}C$, The optimum extraction condition of total vitamin C in fermented persimmon leaf tea was 3min and 5min at $80^{\circ}C\;and\;90^{\circ}C$, 5min and 10min at $60^{\circ}C$. The total vitamin C of conventional persimmon leaf tea was so little that could not be measured by DNP method. The SOD-like activity of conventional and steamed persimmon leaf teas were increased and that of fermented showed the trend of increasing-decreasing-increasing. Fermented persimmon leaf tea had higher SOD-like activity than conventional and steamed at all tested conditions, and the optimum extraction condition of SOD-like activity was 3min and 10min at $80^{\circ}C\;and\;90^{\circ}C$.

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Effects of Temperature Stress and Paraquat on SOD Activity and Photochemical Efficiency of PSII in Leaves of Araliaceae Plants (두릅나무과 식물의 SOD 활성과 광계II의 광화학적 효율에 미치는 온도 스트레스와 Paraquat의 영향)

  • 오순자;고정군;김응식;오문유;고석찬
    • Korean Journal of Environmental Biology
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    • v.17 no.2
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    • pp.199-204
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    • 1999
  • The effects of temperature stress and paraquat on the superoxide dismutase (SOD) activity and the photochemical efficiency of photosystem II were studied in the leaves of Araliaceae plants. The SOD activity of Acanthopanax koreanum leaf discs increased at 4$^{\circ}C$ and 28$^{\circ}C$, and increased significantly at 4$^{\circ}C$ ,28$^{\circ}C$ and 35$^{\circ}C$ in the presence of paraquat. However, the SOD activity of Dendropanax morbifera leaf discs decreased at 4$^{\circ}C$, 28$^{\circ}C$ and 35$^{\circ}C$ regardless of paraquat treatment. The photochemical efficiency of photosystem II, Fv/Fm, of leaf discs of A. koreanum and D. morbifera fell remarkably at 35$^{\circ}C$. In the presence of paraquat, the Ev/Fm values fell slightly at 4$^{\circ}C$ in A. koreanum leaf discs and at 35$^{\circ}C$, in D. morbifera leaf discs. These results indicate that A. koreanum plants are more resistant to temperature stress or oxidative stress than D. morbifera plants although their photochemical efficiency falls slightly at 4$^{\circ}C$ in the presence of paraquat.

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Cloning and Characterization of the Cu,Zn Superoxide Dismutase (SOD1) cDNA from the Spider, Araneus ventricosus

  • Choi Young Soo;Choo Young Moo;Li Jianhong;Sohn Hung Dae;Jin Byung Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.10 no.1
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    • pp.73-77
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    • 2005
  • A Cu,Zn superoxide dismutase (SOD1) cDNA was cloned from the spider, Araneus ventricosus. The A. ventricosus SOD1 (AvSOD1) cDNA contains an open reading frame of 495 bp encoding 165 amino acid polypeptide with a predicted molecular mass of 17,114 Da and pI of 6.55, and possesses the typical metal binding ligands of six histidines and one aspartic acid common to SOD1s. The deduced amino acid sequence of the AvSOD1 cDNA showed $51\%$ identity to Ceratitis capitata SOD1, and $50\%$ to SOD1 sequences of both Drosophila melanogaster and Chymomyza amoena. Northern blot analysis revealed the presence of AvSOD1 transcripts in all tissues examined.

The Study of Superoxide dismutase (SOD) and SOD-mimic Compounds in Panax ginseng C.A.Meyer

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    • Korean Journal of Plant Resources
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    • v.10 no.2
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    • pp.188-193
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    • 1997
  • Panax ginseng C.A.Meyer,1 to 5 years old were electrophored and were stained for SOD activity. The result indicated a total of 13 distinct form of the enzyme and the pattern of achromatic bands were not different according to ages. Nine of the enzyme activities were eliminated with cyanide or peroxide treatment and were resistant to treatment of chloroform plus ethanol. It suggested that they may be cupro-zinc containing SOD, whereas four were cyanide or peroxide resistant and were eliminated with cholroform plus ethanol treatment. They may be manganese containing SOD. Ginseng roos. 1 to 5 years old were analyzed for their SOD measurement of SOD activities of all extracts, the significant difference of SOD activities were not shown according to ages. All ginseng extracts had the total SOD activities of all extracts, the significant difference of SOD activities were not shown according to ages. All ginseng extracts had the total SOD activities of about 700-800 unit/g of fresh weight. Therefore, the SOD activities from SOD-mimic compounds were higher than one from SOD. The ratio between the SOD activity from SOD-mimic compounds and one from true SOD was approximately 2:1 to 3:1.

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Permission-Based Separation of Duty Model on Role-Based Access Control (역할기반 접근제어 환경에서 접근권한 기반의 임무분리 모델)

  • Oh Se-Jong
    • The KIPS Transactions:PartC
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    • v.11C no.6 s.95
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    • pp.725-730
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    • 2004
  • Separation of Duty(SOD), with delegation, is one of important security principles in access control area. The role-based access control model adopts SOD principle, but it has some problems; SOD concept is inconsistent with role hierarchy, permissions that have no relation with SOD may be restricted, and delegation may violate SOD. We propose permission-based SOD model on role-based access control. We establishes SOD as a set of permissions instead of role level SOD. Furthermore we propose a principle of role activation. It solves SOD problems of RBAC and supports easy implementation of SOD policy.