• Title/Summary/Keyword: Cu/ZnSODs

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Copper, Zinc-Superoxide Dismutase (Cu/Zn SOD) Gene During Embryogenesis of Bombyx mori: Molecular Cloning, Characterization and Expression

  • Hong, Sun-Mee;Kang, Seok-Woo;Goo, Tae-Won;Kim, Nam-Soon;Lee, Jin-Sung;Nho, Si-Kab
    • International Journal of Industrial Entomology and Biomaterials
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    • v.13 no.1
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    • pp.23-30
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    • 2006
  • BmCu/Zn SOD was isolated from early embryo of Bombyx mori using microarray analysis. The BmCu/Zn SOD gene was observed during the early embryonic stage with the strongest signal found at the unfertilizaion, fertilization and blastoderm stages. The BmCu/Zn SOD gene encodes a protein of 154 amino acids with a calculated Mr of 15 kDa. The deduced amino acid sequence of BmCu/Zn SOD indicated that the residues that form on the Cu/Zn binding site are conserved and that the sequence is a 60% identity to that of M. domestica. In a phylogenetic tree, Bm SOD was also close to Drosophila SODs rather than other insect SODs. The BmCu/Zn SOD gene exists as a single copy in the genome. Transcripts of BmCu/Zn SOD cDNA were identified by northern blot analysis. The expression of the BmCu/Zn SOD gene was observed weakly in most of larvae, pre-pupae, pupae and adult tissues. Also, the BmCu/Zn SOD gene was observed in early embryonic stage. Although the roles of SODs remains to be further elucidated, the high expression of BmCu/Zn SOD gene at before 24 h post fertilization suggests that this gene is of general importance during early embryogenesis in the Bombyx mod.

Test of Superoxide Dismutase Characteristics and Antioxidant Activity in Perilla Leaves (들깨잎에 함유된 Superoxide Dismutase의 특성 및 항산화 활성 검정)

  • Chung, Ill-Min;Yun, Song-Joong;Kim, Jung-Tae;Gwag, Jae-Gyun;Sung, Jae-Duck;Suh, Hyung-Soo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.4
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    • pp.504-511
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    • 1995
  • This study was conducted to investigate the superoxide dismutase (SOD) characteristics and antioxidant activity by nonenzymatic(Fe$^{2+}$/Ascorbate) and Fe$^{3+}$-ADP/NADPH method in perilla(Perilla frutescens var. japonica Hara.) and jaso(Perilla frutescens Briton var. acuta Kudo.) leaves. The characteristics were evaluated by the nitro blue tetrazolium reduction method. Perilla leaves contained three or four major SODs depending on the varieties. The inhibitor test indicated that the Perilla leaves contained two Cu /ZnSODs and one or two FeSODs, but Jaso leaves have only Cu/ZnSOD. However, no varietal differences were detected in the Cu /ZnSOD isozyme patterns. FeSODs, however, showed different varietal isozyme patterns through the different combinations of the two FeSOD isozymes. Among MeOH extractes, "mil yang 2" showed very strong antioxidant activity. Relatively large differences in the levels of SOD and antioxidant activity detected in the Perilla varietites. There was significantly different in the comparison between perilla leaves and red jaso leaves.s.etween perilla leaves and red jaso leaves.

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Screening of Korean Medicinal and Food Plants with Antioxidant Activity (한국 약용 및 식용식물들의 항산화성 식물탐색)

  • Chung, Il-Min;Kim, Kwang-Ho;Ahn, Joung-Kuk
    • Korean Journal of Medicinal Crop Science
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    • v.6 no.4
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    • pp.311-322
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    • 1998
  • Sixty medicinal and food plants native to Korea were mainly selected with old traditional habit and antioxidant activity was investigated. The 80% EtOH extracts of sixty medicinal and food plants were screened for antioxidant activity. Antioxidant activity was measured by the TBA (Thiobarbituric acid), DPPH (1, 1-Diphenyl-2-picrylhydrazyl), SOD (superoxide dismutase) which was evaluated by the nitro blue tetrazolium(NBT) reduction method. Among sixty plants, black Glycine max(87. 3%) and Solanum nigrum (80.6%) exhibited the highest antioxidant activity by TBA and DPPH methods, respectively. Also, 10 species extracts including black Glycine max showed the high activity value in these two methods. The SOD characteristics on black Glycine max seed extracts which showed the highest SOD activity (53.5%) exhibited four major SODs; two Cu/ZnSODs and two FeSODs. However, Adenophaora vertidllata which showed lowest SOD value (10.4%) had only Cu/Zn SOD. No varietal differences in the high SOD value were detected in the Cu/Zn SOD isozyme patterns.

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Role of Inducibility of Superoxide Dismutases and Metallothionein of Mouse Lungs by Paraquat in Aging (Paraquat에 의한 생쥐 폐의 Superoxide Dismutases와 Metallothionein의 유도능과 노화와의 관계)

  • Lee, Tae-Bum;Park, Yoo-Hwan;Choi, Cheol-Hee
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.5
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    • pp.579-590
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    • 2001
  • Background : The aging process may be induced, at least in part, by reactive oxygen species(ROS). It has been thought that the lung could be a good source of ROS because it has a high oxygen tension. In the present study, we invetigated the inducibility of the first and last lines against oxidative stress, superoxide dismutases(CujZn-SOD and Mn-SOD) as a scavenger of ${O_2}^-\;{\cdot}$ and metallothionein(MT) as a scavenger of $OH{\cdot}$, respectively, in mouse lungs with age. Methods : Oxidative stress was induced by paraquat, an intracellular superoxide generator, at 1, 4, 8, and 12 months of age and then SODs and MT mRNAs were determined by RT-PCR method. Results : The steady-state level of Mn-SOD mRNA increased from 1 to 8 months but decreased thereafter. However, Mn-SOD mRNA was not induced by paraquat after 1 month. On the other hand, there was no change in the steady-state level of Cu/Zn-SOD mRNA, which decreased abruptly at 12 months of age. Additionally, Cu/Zn-SOD mRNA was not induced by paraquat at any age. There was no change in the steady-state level of MT mRNA with age whereas its inducibility by paraquat was intact at all ages. Conclusion : These results indicate that lack of induction of SODs with age may be one of the causative factors in the aging process while induction of MT may play an important role in the defense against oxidative stress. It is therefore implicated that the tissue antioxidant/prooxidant balance could be one of determinants of mean life span.

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Isoform-Specific Responses of Superoxide Dismutase to Oxidative Stresses and Hormones in Parquat-Tolerant Rehmannia glutinosa

  • Jamal, Arshad;Yoo, Nam-Hee;Yun, Song-Joong
    • Journal of Crop Science and Biotechnology
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    • v.10 no.1
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    • pp.8-12
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    • 2007
  • All accessions of Rehmannia glutinosa show the unique characteristic of intrinsic tolerance to paraquat. The higher level of endogenous superoxide dismutase(SOD) activity and its increase upon paraquat treatment indicated the involvement of SOD in the tolerance mechanism to paraquat in R. glutinosa. In this study, we examined the isoform-specific response of SOD to oxidative stresses and hormones. Six SOD isoforms were found in the leaf, and they were identified as two MnSODs(named MnSOD I and MnSOD II, in order of increasing mobility), one FeSOD and three Cu/ZnSODs(named Cu/ZnSOD I, Cu/ZnSOD II, and Cu/ZnSOD III, in order of increasing mobility). MnSOD I, MnSOD II, FeSOD, Cu/ZnSOD I, Cu/ZnSOD II, and Cu/ZnSOD III, contributed to 4, 11, 7, 15, 30, and 32% of the total SOD activity, respectively. Total SOD activity levels in the leaf were increased by 4, 24, and 21% by paraquat, salicylic acid(SA), and yeast extract(YE), respectively, but little by ethephon. Six SOD isoforms responded differentially to these stresses and hormones. The activities of all the isoforms were increased by YE and SA except that of MnSOD I which was decreased by SA. The activities of MnSOD I, FeSOD, and CuZnSOD I were increased by paraquat. These results suggest that amelioration of oxidative stresses by SOD is fine-tuned by the differential expression of isoforms in R. glutinosa.

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Isolation and Transcriptional Expression of CuZn Superoxide Dismutase from Codonopsis lanceolata

  • Lee, Kang;In, Jun-Gyo;Yu, Chang-Yeon;Yun, Song-Joong;Min, Byung-Hoon;Rho, Yeong-Deok;Kim, Moo-Sung;Yang, Deok-Chun
    • Plant Resources
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    • v.7 no.3
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    • pp.163-169
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    • 2004
  • To investigate the defense mechanism against the abiotic stress, a cDNA clone encoding a CuZn superoxide dismutase (CuZnSOD) protein was isolated from a cDNA library prepared from tabroot mRNAs of Codonopsis lanceolata. The eDNA, designated ClSODCc, is 799 nucleotides long and has an open reading frame of 459 bp with a deduced amino acid sequence of 152 residues. The deduced amino acid sequence of ClSODCc matched to the previously reported CuZnSODs. Consensus amino acid residues (His-45, -47, -62, -70, -79, -119 and Asp-82) were involved in Cu-, Cu/Zn-, and Zn- binding ligands. The deduced amino acid sequence of ClSODCc showed high homologies (82%-86%) regardless of species. Expression of ClSODCc by oxidative stress was increased up to 1 h after treatment and declined gradually. Much earlier and stronger expression of ClSODCc was observed in the cold stress treatment.

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Expression of a Cu-Zn Superoxide Dismutase Gene in Response to Stresses and Phytohormones in Rehmannia Glutinosa

  • Park, Myoung-Ryoul;Ryu, Sang-Soo;Yoo, Nam-Hee;Yu, Chang-Yeon;Yun, Song-Joong
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.5
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    • pp.270-275
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    • 2005
  • Superoxide dismutases (SOD) are metalloenzymes that convert $O_2^-\;to\;H_2O_2$. Rehmannia glutinosa is highly tolerant to paraquat-induced oxidative stress. The primary objective of this study was to characterize regulation of SOD gene expression in R. glutinosa in response to oxidative stresses and hormones. A full-length putative SOD clone (RgCu-ZnSOD1) was isolated from the leaf cDNA library of R. glutinosa using an expressed sequence tag clone as a probe. RgCu-ZnSOD1 cDNA is 777 bp in length and contains an open reading frame for a polypeptide consisted of 152 amino acid residues. The deduced amino acid sequence of the clone shows highest sequence similarity to the cytosolic Cu-ZnSODs. The two to three major bands with several minor ones on the Southern blots indicate that RgCu-ZnSOD1 is a member of a small multi-gene family. RgCuZnSOD1 mRNA was constitutively expressed in the leaf, flower and root. The expression of RgCu-ZnSOD1 mRNA was increased about 20% by wounding and paraquat, but decreased over 50% by ethylene and $GA_3$. This result indicates that the RgCu-ZnSOD1 expression is regulated differentially by different stresses and phytohormones at the transcription level. The RgCu-ZnSOD1 sequence and information on its regulation will be useful in investigating the role of SOD in the paraquat tolerance of R. glutinosa.

Molecular Cloning and Expression of a Cu/Zn-Containing Superoxide Dismutase from Thellungiella halophila

  • Xu, Xiaojing;Zhou, Yijun;Wei, Shanjun;Ren, Dongtao;Yang, Min;Bu, Huahu;Kang, Mingming;Wang, Junli;Feng, Jinchao
    • Molecules and Cells
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    • v.27 no.4
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    • pp.423-428
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    • 2009
  • Superoxide dismutases (SODs) constitute the first line of cellular defense against oxidative stress in plants. SODs generally occur in three different forms with Cu/Zn, Fe, or Mn as prosthetic metals. We cloned the full-length cDNA of the Thellungiella halophila Cu/Zn-SOD gene ThCSD using degenerate RT-PCR and rapid amplification of cDNA ends (RACE). Sequence analysis indicated that the ThCSD gene (GenBank accession number EF405867) had an open reading frame of 456 bp. The deduced 152-amino acid polypeptide had a predicted molecular weight of 15.1 kDa, an estimated pI of 5.4, and a putative Cu/Zn-binding site. Recombinant ThCSD protein was expressed in Escherichia coli and assayed for SOD enzymatic activity in a native polyacrylamide gel. The SOD activity of ThCSD was inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide, confirming that ThCSD is a Cu/Zn-SOD. Northern blotting demonstrated that ThCSD is expressed in roots, stems, and leaves. ThCSD mRNA levels increased by about 30-fold when plants were treated with sodium chloride (NaCl), abscisic acid (ABA), and indole-acetic acid (IAA) and by about 50-fold when treated with UVB light. These results indicate that ThCSD is involved in physiological pathways activated by a variety of environmental conditions.

Antiproliferative Effect of Artemisia argyi Extract against J774A.1 Cells and Subcellular Superoxide Dismutase (SOD) Activity Changes

  • Lee, Tea-Eun;Park, Sie-Won;Min, Tae-Jin
    • BMB Reports
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    • v.32 no.6
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    • pp.585-593
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    • 1999
  • The water and methanol extracts of Artemisia argyi showed significant cytotoxicities against J774A.1 cells but not so much against normal leukocytes. The cytotoxicities were found to be dependent on the extract concentration and the incubation time. The concentration of water and methanol extracts inhibiting 50% of cell proliferation ($IC_{50}$) were estimated to be 44.2 mg/ml and 71.6 mg/ml, respectively. In the presence of Artemisia argyi water extract, total superoxide dismutase (CuZnSOD and MnSOD) activities of media, cytoplasmic and mitochondrial fractions of J774A.1 cells increased in accordance with cytotoxicity. MnSOD was found to be the main component of enhanced total SOD activities, particulary in the mitochondrial fraction. In contrast to SOD, catalase and glutathione peroxidase (GPx) were not found in any instance of the current investigation. In addition, substantial amount of $O_2^-$ appeared to be generated in the mitochondrial fraction under the influence of Artemisia argyi. All data put together, it is postulated that Artemisia argyi extracts seem to stimulate $O_2^-$ generation in mitochondria of J774A.1 cells with concomitant increases of SODs. Since $H_2O_2$, the reaction product of SOD on $O_2^-$, is known to be readily converted to very toxic $OH{\cdot}$ in the absence of catalase and/or GPx cooperation, toxicity derived from ROS such as $O_2^-$, $H_2O_2$, and $OH{\cdot}$ may be the main cause of necrosis and/or apoptosis of J774A.1 cells.

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A Study of the Generation of Transgenic Chickens That Express Human SOD-3 Protein (사람의 SOD-3 단백질을 발현하는 형질전환 닭 생산 연구)

  • Byun, S.J.;Park, C.;Kim, J.A.;Woo, J.S.;Lee, H.C.;Kim, T.Y.;Kim, S.H.;Seong, H.H.;Park, J.K.;Jeon, I.S.
    • Korean Journal of Poultry Science
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    • v.35 no.3
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    • pp.241-245
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    • 2008
  • Lentiviral vector system is efficient vehicles for the delivery of exogenous genes, and it is generally used in the generation of transgenic chickens. In this study, we used recombinant lentiviral vectors to generate transgenic chicks that express the human superoxide dismutase-3 gene driven by the chicken ovalbumin promoter. It is well known that superoxide dismutases(SODs) are believed to play a crucial role in protecting cells against oxygen toxicity. There are three forms of SOD proteins: cytosolic Cu-Zn SOD, mitochondrial Mn SOD, and extracellular SOD(SOD-3). The recombinant lentivirus containing the human SOD-3 gene was injected into the subgerminal cavity of freshly laid eggs. Subsequently, the embryos were incubated to hatch using phases II and III of the surrogate shell ex vivo culture system. From 341 injected embryos, the 78 chicks hatched after 21 days incubation. The hatched chicks were screened for the human SOD-3 gene by using PCR. Two of 47 male chickens that survived to sexual maturity contained the human SOD-3 gene in their semen. These results showed that our transgenic chicken generation system was completely established.