• 제목/요약/키워드: Superoxide Dismutase Gene Expression

검색결과 136건 처리시간 0.036초

녹차가 전자파 조사 흰쥐 간조직의 Superoxide Dismutase 및 Glutathione Peroxidase 유전자 발현에 미치는 영향 (Effect of Green Tea on Gene Expression of Superoxide Dismutase and Glutathione Peroxidase in Rat Liver Exposed to Microwaves)

  • 최정화
    • Journal of Nutrition and Health
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    • 제33권7호
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    • pp.733-738
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    • 2000
  • The purpose of this study was to investigate the effects of green tea on gene expression of superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) in rat liver exposed to microwave. Sprague-Dawley male rats with 200$\pm$10g body weight were assigned to normal and microwave exposed groups : microwave exposed groups ; microwave exposed groups were divided two groups : microwave(MW) group which was administrated the distilled water and green tea(GT) group which was administrated the green tea extracts. The rats were irradiated with microwave at frequence of 2.45 GHz for 15 min and then the gene expression in the damaged tissue were investigated at 0.1, 3, 4,6 and 8 days after the microwave irradition to compared with the normal group. The level of SOD gene expression in MW group was lower than the normal group within 6 days but that of GT group as higher than MW group. These results may imply that green tea stimulates SOD expression and there by protecting tissues from free radicals. The GSH-Px gene was expressed a little bit lower than the normal group but that of GT group was expressed to higher lever than MW group from 4 days after irradiation. These results suggest that the administration of green tea extract may activate antioxidative gene expressions such as SOD and GSH-Px in rat and that may help to recover liver tissues from microwave damage by removing hazardous free radicals and oxidized by products from cells.

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Cloning, DNA Sequence Determination, and Analysis of Growth-Associated Expression of the sodF Gene Coding for Fe- and Zn-Containing Superoxide Dismutase of Streptomyces griseus

  • Kim, Ju-Sim;Lee, Jeong-Kug
    • Journal of Microbiology and Biotechnology
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    • 제10권5호
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    • pp.700-706
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    • 2000
  • Iron- and zinc-containing superoxide dismutase (FeZnSOD) and nickel-containing superoxide dismutase (NiSOD) are cytoplamic enzymes in Streptomyces griseus. The sodF gene coding for FeZnSOD was cloned from genomic Southern hybridization analysis with a 0.5-kb DNA probe, which was PCR-amplified with facing primers corresponding to the N-terminal amino acid of the purified FeZnSOD of S. griseus and a C-terminal region which is conserved among bacterial FeSODs and MnSODs. The sodF open reading frame (ORF) was comprised of 213 amino acid (22,430 Da), and the deduced sequence of the protein was highly homologous (86% identity) to that of FeZnSOD of Streptomyces coelicolor. The FeZnSOD expression of exponentially growing S. griseus cell was approximately doubled as the cell growth reached the early stationary phase. The growth-associated expression of FeZnSOD was mainly controlled at the transcriptional level, and the regulation was exerted through the 110 bp regulatory DNA upstream from the ATG initiation codon of the sodF gene.

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Effects of Gene Expression of Photobacterium leiognathi CuZn Superoside Dismutase (PSOD) by lacZ Promotor Control under Oxidative Stress

  • Kim, Young-Gon
    • 미생물학회지
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    • 제30권6호
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    • pp.460-465
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    • 1992
  • The effect of PSOD expression on lacZ-sodP fusion (pYK4) was explored in Escherichia coli sodA sodB mutants (QC774) under oxidative stress. In this system, although .betha.-galactosidase activity was not fully induced by isopropyl-1-thio-.betha.-galactosidase (IPTG) and was inhibited by glucose, functional PSOD was under lacZ promotor control and was induced by IPTC, lactose, PQ and copper isons, finally, the results show that higher PSOD expression leel was consistently importnat in defending against superoxide radicals.

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PAH를 분해할 수 있는 Pseudomonas rhodesiae KK1의 SOD 유전자의 동정 및 분자학적 특성 분석 (Identification and Molecular Characterization of Superoxide Dismutase Genes in Pseudomonas rhodesiae KK1 Capable of Polycyclic Aromatic Hydrocarbon Degradation)

  • 이동헌;오계헌;김승일;강형일
    • 생명과학회지
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    • 제26권1호
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    • pp.75-82
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    • 2016
  • Pseudomonas rhodesiae KK1은 이미 주요한 환경오염물질인 anthracene, naphthalene, phenanthrene과 같은 다환성 방향족 화합물(PAHs)을 분해할 수 있음을 보고한 바 있다. 흥미롭게도, superoxide dismutase를 비롯한 항산화 유전자는 환경오염물질에 반응하여 다른 수준으로 발현됨이 알려져 있다. 본 연구는 균주 KK1에서 PAHs 분해에 간접적으로 관계될 것으로 여겨지는 superoxide dismutase 유전자의 존재를 동정하고 세 가지 PAHs를 기질로 하여 생장한 세포에서 superoxide dismutase 유전자의 발현 양상을 조사하고자 수행하였다. P. rhodesiae KK1에서 항산화 기작에 관여하는 두 가지지 형의 superoxide dismutase인 Mn-superoxide dismutase (sodA)와 Fe-superoxide dismutase (sodB) 유전자를 동정하고 그 특성을 규명하였다. 균주 KK1에서 발견된 sodA 유전자는 141개의 아미노산 유전자를 기준으로 P. fluorescens Pf-5의 Mn-sod와 95%, sodB 유전자는 135개 아미노산을 기준으로 P. fluorescens Pf-5의 Fe-sod와 99%의 가장 높은 상동성을 나타내었다. sod 유전자 단편을 탐침자로 사용한 Southern 혼성화 반응 결과 적어도 두 개 이상의 superoxide dismutase 유전자가 균주 KK1에 존재함을 규명하였다. RT-PCR 분석을 통해 sodA 및 sodB 유전자들은 anthracene보다 naphthalene과 phenanthrene에 반응하여 더 강하게 발현함을 보여주었다. 포도당과 PAHs를 기질로 사용하여 생장한 세포에서 sodA와 sodB 유전자는 활성 상태로 존재함이 밝혀졌다.

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

  • Baek, Kwang-Hyun;Skinner, Daniel Z.
    • 한국환경농학회지
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    • 제29권1호
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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.

사람의 세포질 Superoxide Dismutase 유전자의 클로닝과 대장균내에서의 대량발현에 관한 연구 (Molecular Cloning and High-Level Expression of Human Cytoplasmic Superoxide Dismutase Gene in Escherichia coli)

  • 이우길;김영호;양중익;노현모
    • 미생물학회지
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    • 제28권2호
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    • pp.91-97
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    • 1990
  • 생체내의 유해산소를 제거하는 superoxide dismutase (superoxide : superoxide oxidoreductase E.C.1.15.1.1) 중 세포질내에서 그 활성을 지니는 인체의 세포질 superoxide dismuta~ie (SODl) 유전자를 사람의 간 cDNA library로부터 동위원소로 표지된 oligonucleotide probe를 이용, in situ plaque hybridization 방법으로 선별 분리하여 내장균 벡터로 클로닝하였다. 이 클론은 SOD1 유전자의 5"L"TR과 3’UTR을 포함한 1.6 kb 정도의 cDNA였다 SOD1 구조유전자만을 선택적으로 분리하기 위해서 ATG를 포함하는 sense strand primer와 3’UTR 부위의 antisense strand primer를 이용하여 중합효소연쇄반응(Polymerase Chain Reaction) 방법을 써서 SOD1 구조유전자 부위만을 선택적으로 증폭시켰다. Taq DNA polymerase에 의해 증폭된 DNA를 벡터 pUCl9의 multiple cloning site (MCS) 내의 Hinc II 위치에 넣였으며 이 insert DNA를 M13 mp19으로 옮겨 dideoxy chain termination 방법으로 sequenase를 사용하여 염기서열을 결정하였다. 클론닝된 cDNA는 153개의 아미노산을 포함하고 있는 하나의 open reading frame (ORF)을 가셨다. 중합효소연쇄반응에 의해 이때 증폭된 SOD1 구조유전자를 $\lambda P_{L}$ 프로모터를 포함하고 있는 발현 벡터 pUPL에 옮긴 후 대장균에서 대량으로 발현시켰다. 이때 발현된 단백질 SOD1은 고유의 효소활성을 가지고 있었다.

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Expression and Purification of Recombinant Superoxide Dismutase (PaSOD) from Psychromonas arctica in Escherichia coli

  • Na, Ju-Mee;Im, Ha-Na;Lee, Kyung-Hee
    • Bulletin of the Korean Chemical Society
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    • 제32권7호
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    • pp.2405-2409
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    • 2011
  • The psychrophilic bacteria Psychromonas arctica survives at subzero temperatures by having adapted several protective mechanisms against freezing and oxidative stresses. Many reactive oxygen species are likely generated in P. arctica as a result of reduced metabolic turnover rates. A previous study identified the pasod gene for superoxide dismutase from P. arctica using a series of PCR amplifications. Here, upon cloning into a His-tag fused plasmid, the sod gene from P. arctica (pasod) was successfully expressed by IPTG induction. His-tagged PaSOD was subsequently purified by $Ni^{2+}$-NTA affinity chromatography. The purified PaSOD exhibited a higher SOD activity than that of Escherichia coli (EcSOD) at all temperatures. The difference in activity between PaSOD and EcSOD becomes even more significant at 4$^{\circ}C$, indicating that PaSOD plays a functional role in the cold adaptation of P. arctica in the Arctic.

Transcriptional Activation of CuIZn Superoxide Dismutase And Catalase Genes by Panaxadiol Ginsenosides Extracted From Panax ginseng

  • Chang, Mun-Seog;Yoo, Hae-Yong;Rho, Hyune-Mo
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1998년도 Advances in Ginseng Research - Proceedings of the 7th International Symposium on Ginseng -
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    • pp.63-70
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    • 1998
  • Superoxide dismutase (SOD) and catalase constitute the first coordinated unit of defense against reactive oxygen species. Here, we examined the effect of ginseng saponins on the induction of SOD and catalase gene expression. To explore this possibility, the upstream regulatory promoter region of Cu/Zn superoxide dismutase (SODI) and catalase genes were linked to the chloramphenicol acetyl-transferase (CATI structural gene and introduced into human hepatoma HepG2 cells. Total saponin and panaxatriol did not activate the transcription of SODI and catalase genes but panaxadiol increased the transcription of these genes about 2-3 fold. Among the Panaxadiol ginsenosides, the Rb2 subtraction appeared to is a major induce of SODI and catalase genes. Using the deletion analyses and mobility shift assays, we showed that the 5051 gene was greatly activated by ginsenoside Rba through transcription factor AP2 binding sites and its induction. We also examined the effect of the content ratio of panaxadiol extracted from various compartment of ginseng on the transcription of 5031 gene. Saponin extract that contains 2.6-fold more PD than PT from the fine root Increased the SODI induction about 3-fold. These results suggest that the panaxadiol fraction and its ginsenosides could induce the antioxidant enzymes, which are important for maintaining cell viability by lowering level of oxygen radical generated from intracellular metabolism.

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Saccharomyces cerevisiae에서 이온화 방사선과 N-acetyl-L-cysteine 처리에 따른 세포 생존과 Superoxide Dismutase와 Catalase 유전자 발현 (Cell Survival and Expression of Superoxide Dismutase and Catalase Genes in Saccharomyces cerevisiae Treated with N-acetyl-L-cysteine and Ionizing Radiation)

  • 박지영;백동원;모하마드닐리;김진규
    • 환경생물
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    • 제29권1호
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    • pp.61-67
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    • 2011
  • NAC는 GSH의 전구물질로, thiol기를 포함하는 항산화제 중 하나로 잘 알려져 있으며, 방사선 조사 시 발생하는 생체 내 영향을 감소시켜 생체 손상의 방호 및 회복에 도움을 주는 방사선 방어제로 이용된다. S. cerevisiae에서 항산화제 NAC를 전처리 함에 따라 이온화 방사선 조사에 따른 효모의 세포사멸 방어효과 및 superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx)와 같은 항산화 효소들의 유전자 발현을 분석하여 NAC의 항산화적 효과를 확인하였다. 효모는 다양한 농도의 NAC 전처리 후 다양한 선량의 이온화 방사선에 조사되었으며, 세포생존율은 세포형성단위(CFU)를 계수해 측정되었고, 항산화 효소의 유전자 발현은 real-time PCR수행 후 분석하였다. 우선적으로 효모에 NAC 처리를 위한 적정농도를 확인하였는데, 35 mM 이상의 NAC 농도에서 효모세포의 성장이 억제 되었다. NAC 전처리는 감마선 조사에 의한 세포사멸을 방어하지 않았으며, 100 Gy 방사선 조사는 항산화 효소들의 유전자 발현을 유도하였다. NAC 전처리 후 항산화 효소들의 유전자 발현은NAC의 농도 증가에 따라 감소하였다. 이러한 결과로,NAC의 높은 농도(35 mM 이상)는 효모세포의 성장을 저해하며, NAC는 이온화 방사선 조사에 따른 세포사멸을 방어할 수 없으나, 생체 내에서 활성산소종을 제거 하여 세포를 보호하는 유용한 항산화제임을 알 수 있었다.

Extracellular Superoxide Dismutase (EC-SOD) Transgenic Mice: Possible Animal Model for Various Skin Changes

  • Kim, Sung-Hyun;Kim, Myoung-Ok;Lee, Sang-Gyu;Ryoo, Zae-Young
    • Reproductive and Developmental Biology
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    • 제30권4호
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    • pp.229-234
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    • 2006
  • We have generated transgenic mice that expressed mouse extracellular superoxide dismutase (EC-SOD) in their skin. In particular, the expression plasmid DNA containing human keratin K14 promoter was used to direct the keratinocyte-specific transcription of the transgene. To compare intron-dependent and intron-independent gene expression, we constructed two vectors. The vector B, which contains the rabbit -globin intron 2, was not effective for mouse EC-SOD overexpression. The EC-SOD transcript was detected in the skin, as determined by Northern blot analysis. Furthermore, EC-SOD protein was detected in the skin tissue, as demonstrated by Western blot analysis. To evaluate the expression levels of EC-SOD in various tissues, we purified EC-SOD from the skin, lungs, brain, kidneys, livers, and spleen of transgenic mice and measured its activities. EC-SOD activities in the transgenic mice skin were approximately 7 fold higher than in wild-type mice. These results suggest that the mouse overexpressing vector not only induces keratinocyte-specific expression of EC-SOD, but also expresses successfully functional EC-SOD. Thus, these transgenic mice appeared to be useful for the expression of the EC-SOD gene and subsequent analysis of various skin changes, such as erythema, inflamation, photoaging, and skin tumors.