• 제목/요약/키워드: sod gene

검색결과 226건 처리시간 0.021초

Cloning, Expression, and Characterization of Superoxide dismutase from Aquifex Pyophilus, a Hyperthermophilic Bacteria

  • Rhim, Jae-Hwan;Yesun Han;Kim, Sung-Hou;Yunje Cho
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 1996년도 정기총회 및 학술발표회
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    • pp.30-30
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    • 1996
  • A suproxide dismutase gene of Aquifex pyroprolus, a novel marine hypenhermophilic bacterium, was cloned, expressed, and characterized. The SOD of A pyrophilus (ApSOD) is an iron-containing homo-oligomeric protein with a monomeric molecular weight of 24.2 kDa. the amino acid sequence is similar to those of known Mn- and Fe-SODs from thermophilic archaea, and metal binding residues in all SOD sequences from different species are also conserved in A. pyrophilus SOD. (omitted)

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Anti-oxidative Effect of Piperine from Piper nigrum L. in Caenorhabditis elegans

  • Park, Hyun Mee;Kim, Jun Hyeong;Kim, Dae Keun
    • Natural Product Sciences
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    • 제25권3호
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    • pp.255-260
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    • 2019
  • Piper nigrum L. (Piperaceae), which is a well-known food seasoning, has been used as a traditional medicine for the treatment of vomiting, abdominal pain, diarrhea and anorexia in Korea, China and Japan. Methanol extract from the fruit of P. nigrum was successively partitioned as n-hexane, methylene chloride, ethyl acetate, n-butanol and $H_2O$ soluble fractions. Among those fractions the ethyl acetate soluble fraction showed the most potent DPPH radical scavenging activity, and piperine was isolated from the ethyl acetate fraction. To know the antioxidant activity of piperine, we tested the activities of superoxide dismutase (SOD) and catalase together with oxidative stress tolerance and intracellular ROS level in Caenorhabditis elegans. To investigate whether piperine-mediated increased stress tolerance was due to regulation of stress-response gene, we quantified SOD-3 expression using transgenic strain including CF1553. Consequently, piperine enhanced SOD and catalase activities of C. elegans, and reduced intracellular ROS accumulation in a dose-dependent manner. Moreover, piperine-treated CF1553 worms exhibited significantly higher SOD-3::GFP intensity.

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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계지복령환이 Cholesterol 식이 생쥐의 SOD 유전자발현능에 미치는 영향 (Effects of geagibokrounghwan on superoxide dismutase gene expression in experimental mouse fed cholesterol)

  • 홍윤표;노승현
    • 대한한의학방제학회지
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    • 제6권1호
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    • pp.215-226
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    • 1998
  • Geagibokrounghwan (桂技茯笭湯) has long been used to cure human diseases such as vascular and blood disorders. However, it is still unkown on its action mechanism, physiolosical and biochemical meaning. Thus, many attempts were tried to show the scientific background covering the above mentioned mechanism. The effect of Geagibokrounghwan, which was known to date, as follow; effective circulation of body blood system, proliferation of leucocytes and antioxidative action. In this study, we have applied the Geagibokrounghwan administration and feed to mouse, to see effects on the expression of superoxide dismutase(SOD) mRNA as antioxidative agent and oxygen radical scavenger. Total RNAs includingmRNA have been isolated from liver and white blood cells after mice were fed with cholesterol in high dose. Also, in a separate group, the cholesterol-administrated mice were fed with Geagibokrounghwan to see the effects on SOD transcription. and then reverse transcriptase-polymerase chain reaction (RT-PCR) usion each primer set (SOD-F;GATGAAAGCGGTGT-3'; SOD-R; 5'-CCTGTGGAGTGATT-3') were performed to trace theamounts of mRNA. SOD mRNA was specifically expressed in Geagibokrounghwan-fed mice at 2 weeks after treatment, however, gradually reduced after 4 weeks. These results indicate that Geagibokrounghwan is highly applicable in treatment of the above mentioned human diseases.

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Study of Functional Verification to Abiotic Stress through Antioxidant Gene Transformation of Pyropia yezoensis (Bangiales, Rhodophyta) APX and MnSOD in Chlamydomonas

  • Lee, Hak-Jyung;Yang, Ho yeon;Choi, Jong-il
    • Journal of Microbiology and Biotechnology
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    • 제28권7호
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    • pp.1217-1224
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    • 2018
  • Seaweeds produce antioxidants to counteract environmental stresses, and these antioxidant genes are regarded as important defense strategies for marine algae. In this study, the expression of Pyropia yezoensis (Bangiales, Rhodophyta) ascorbate peroxidase (PyAPX) and manganese-superoxide dismutase (PyMnSOD) was examined by qRT-PCR in P. yezoensis blades under abiotic stress conditions. Furthermore, the functional relevance of these genes was explored by overexpressing them in Chlamydomonas. A comparison of the different expression levels of PyAPX and PyMnSOD after exposure to each stress revealed that both genes were induced by high salt and UVB exposure, being increased approximately 3-fold after 12 h. The expression of the PyAPX and PyMnSOD genes also increased following exposure to $H_2O_2$. When these two genes were overexpressed in Chlamydomonas, the cells had a higher growth rate than control cells under conditions of hydrogen peroxide-induced oxidative stress, increased salinity, and UV exposure. These data suggest that Chlamydomonas is a suitable model for studying the function of stress genes, and that PyAPX and PyMnSOD genes are involved in the adaptation and defense against stresses that alter metabolism.

Development and Characterization of Expression Vectors for Corynebacterium glutamicum

  • Lee, Jinho
    • Journal of Microbiology and Biotechnology
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    • 제24권1호
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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.

마카 추출액의 생리활성 효과 (Biological Activities of Maca (Lepidium meyenii) Extracts)

  • 권윤숙;전인숙;황진현;임동민;강용수;정해정
    • 한국식품영양과학회지
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    • 제38권7호
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    • pp.817-823
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    • 2009
  • 본 실험에서는 마카를 기능성식품 소재로 사용하기 위한 가능성을 알아보기 위하여 마카분말을 다양한 용매로 추출하고 추출수율, 총 폴리페놀 함량, 전자공여능, SOD 유사활성 및 아질산염 소거능 등을 측정하여 최적의 추출조건을 확립하고자 하였다. 일반성분 분석 결과 수분, 조단백질, 조지방, 회분 및 탄수화물 함량은 각각 6.57%, 12.83%, 1.05%, 4.80 및 74.75%로 나타났다. 추출수율은 물을 사용할 경우 46.2%로 메탄올, 에탄올, 아세톤 등으로 추출하는 것보다 약 $2{\sim}200$배 높게 나타났다. 추출용매에 따른 총 폴리페놀 함량은 물 추출액에서 328.68 mg/100 mL로 가장 높게 나타났다. 전자공여능은 메탄올 추출액이 52.36%로 가장 높았고 아질산염 소거능은 pH 1.2의 조건에서 물 추출액이 77.51%로 가장 높게 나타났다. 이상의 결과를 종합하여 최적의 추출용매로 물을 선정하였고 물 추출 온도(30, 70, $100^{\circ}C$)와 시간(1, 3, 5시간)을 달리하여 생리활성 물질의 함량변화를 측정한 결과 $100^{\circ}C$에서 3시간 또는 5시간 추출할 때 총 폴리페놀함량, 전자공여능 및 SOD 유사활성이 다른 조건에서보다 우수한 것으로 나타났다. 따라서 시간과 에너지의 효율적 측면을 고려하여 마카분말을 $100^{\circ}C$에서 3시간 물로 추출하는 것이 가장 적합할 것으로 판단된다.

Involvement of Caenohabditis elegans MAPK Signaling Pathways in Oxidative Stress Response Induced by Silver Nanoparticles Exposure

  • Roh, Ji-Yeon;Eom, Hyun-Jeong;Choi, Jin-Hee
    • Toxicological Research
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    • 제28권1호
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    • pp.19-24
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    • 2012
  • In the present study, toxicity of silver nanoparticles (AgNPs) was investigated in the nematode, Caenohabditis elegans focusing on the upstream signaling pathway responsible for regulating oxidative stress, such as mitogen-activated protein kinase (MAPK) cascades. Formation of reactive oxygen species (ROS) was observed in AgNPs exposed C.elegans, suggesting oxidative stress as an important mechanism in the toxicity of AgNPs towards C. elegans. Expression of genes in MAPK signaling pathways increased by AgNPs exposure in less than 2-fold compared to the control in wildtype C.elegans, however, those were increased dramatically in sod-3 (gk235) mutant after 48 h exposure of AgNPs (i.e. 4-fold for jnk-1 and mpk-2; 6-fold for nsy-1, sek-1, and pmk-1, and 10-fold for jkk-1). These results on the expression of oxidative stress response genes suggest that sod-3 gene expression appears to be dependent on p38 MAPK activation. The high expressions of the pmk-1 gene 48 h exposure to AgNPs in the sod-3 (gk235) mutant can also be interpreted as compensatory mechanisms in the absence of important stress response genes. Overall results suggest that MAPK-based integrated stress signaling network seems to be involved in defense to AgNPs exposure in C.elegans.

Bidirectional Regulation of Manganese Superoxide Dismutase (MnSOD) on the Radiosensitivity of Esophageal Cancer Cells

  • Sun, Guo-Gui;Hu, Wan-Ning;Wang, Ya-Di;Yang, Cong-Rong;Lu, Yi-Fang
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권7호
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    • pp.3015-3023
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    • 2012
  • The mitochondrial antioxidant protein manganese superoxide dismutase (MnSOD) may represent a new type of tumor suppressor protein. Overexpression of the cDNA of this gene by plasmid or recombinant lentiviral transfection in various types of cancer leads to growth suppression both in vitro and in vivo. We previously determined that changes in MnSOD expression had bidirectional effects on adriamycin (ADR) when combined with nitric oxide (NO). Radiation induces free radicals in a manner similar to ADR, so we speculated that MnSOD combined with NO would also have a bidirectional effect on cellular radiosensitivity. To examine this hypothesis, TE-1 human esophageal squamous carcinoma cells were stably transfected using lipofectamine with a pLenti6-DEST plasmid containing human MnSOD cDNA at moderate to high overexpression levels or with no MnSOD insert. Blastidicin-resistant colonies were isolated, grown, and maintained in culture. We found that moderate overexpression of MnSOD decreased growth rates, plating efficiency, and increased apoptosis. However, high overexpression increased growth rates, plating efficiency, and decreased apoptosis. When combined with NO, moderate overexpression of MnSOD increased the radiosensitivity of esophageal cancer cells, whereas high MnSOD overexpression had the opposite effect. This finding suggests a potential new method to kill certain radioresistant tumors and to provide radioresistance to normal cells.