• Journal of Plant Biotechnology
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• v.36 no.2
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• pp.144-148
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• 2009

The transfer of Mn-Superoxide Dismutase (SOD2) gene, complex gene (SA) of CuZnSOD and ascorbate peroxidase (APX), and NDP kinase 2 (NDPK2) gene into Korean 4 cultivars (cvs. Millenium White, Glory Blue, Glory Red, and Glory Purple) and 15 purebred lines of petunia was conducted using Agrobaterium-mediated technique. Two (Wongyo A2-16 and A2-36) of 15 purebred lines and one (cv. Glory Red) of 4 cultivars were effective for the transfer of SOD2 gene. The putative transgenic plants survived on the 2nd selection medium were 124. From PCR analysis, 118 (derived from 4 cultivars and 2 purebred lines) of 124 plants were confirmed to contain marker (npt II ) gene, while 58 of 118 plants did not have target genes. There were no plants with both npt II and SA genes. Twenty seven of 28 SOD2 transgenic plants were re-confirmed as transformants by Sothern analysis. SOD2 and NDPK2 genes were expressed in the transgenic petunias as the ratio of 77.8 to 100.0 % and 23.5%, respectively. T1 seeds were obtained from 36 acclimated transgenic plants (SOD2 34 plus NDPK2) in a glasshouse by self-pollination.

• Journal of Applied Biological Chemistry
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• v.49 no.1
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• pp.1-7
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• 2006

Differential display reverse transcription PCR (DDRT-PCR) analysis was performed on lily 'Marcopolo' bulb scale for isolation of expressed genes during bulblet formation. Cu/Zn lily-superoxide dismutase (LSOD) of 872 bp gene, with ability to scavenge reactive oxygen in stress environment, was isolated. Northern blot analysis showed expression levels of LSOD maximized 12 days after bulblet formation. Ti plasmid vectors were constructed with sense and antisense expressions of LSOD gene and transformed into potato. Southern blot analysis of transgenic potatoes revealed different copies of T-DNA were incorporated into potato genome. In transgenic potatoes, lily SOD gene was overexpressed in sense lines and not in antisense lines. In native polyacrylamide gel electrophoresis analysis, additional engineered LSOD was detected in sense overexpressed transgenic line only. Transgenic potatoes were subjected to oxidative stress, such as herbicide methyl viologen (MV). Transgenic potato lines with sense orientation exhibited increased tolerance to MV, whereas in antisense lines exhibited decreased tolerance. In vitro tuberization of transgenic potato with sense orientation was promoted, but was inhibited in transgenic potato with antisense orientation.

• Journal of Microbiology
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• v.41 no.1
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• pp.28-33
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• 2003

Pycnogenol (PYC) is believed to have potential as a therapeutic agent against free radical-mediated oxidative stress. It is important, therefore, to understand the interactions between PYC and cellular defenses against oxidative stress. Toward this end, we analyzed the survival rates on the gene expression responses of E. coli sod katG mutants to PYC after pre-treatment of PQ or H$_2$O$_2$-mediated stress under aerobic conditions. We identified SOD induced by PYC, but not HP1 in sod hate mutants. A striking result was the PYC induction of SOD with antioxidant property in single katG mutant cells, particularly MnSOD and CuZnSOD. These inductions were further increased with oxidative stress, while HP1 was not induced in these conditions. The effects of pycnogenol treatment on these cells depend in part on its concentration on the stress response. Protective effects of PYC exposure which affected gene expression in cells were consistent with cell survival rates. Our results demonstrate that pycnogenol may alter the stress response gene expression in a specific manner such as SOXRS because PYC induction of single mutant only worked under increased PQ stress. All together our data indicate that SOD activity is essential for the cellular defense against PQ-mediated oxidative stress, suggesting that PYC may not be effective as an antioxidant in only oxidative stress conditions. On the other hand, it was expected that PYC may play a role as a pro-oxidant and if it is available for use, it should be evaluated carefully.

• Molecules and Cells
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• v.19 no.2
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• pp.191-197
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• 2005

The human immunodeficiency virus type 1 (HIV-1) Tat protein transduction domain (PTD) is responsible for highly efficient protein transduction across plasma membranes. In a previous study, we showed that Tat-Cu,Zn-superoxide dismutase (Tat-SOD) can be directly transduced into mammalian cells across the lipid membrane barrier. In this study, we fused the human SOD gene with a Tat PTD transduction vector at its N- and/or C-terminus. The fusion proteins (Tat-SOD, SOD-Tat, Tat-SOD-Tat) were purified from Escherichia coli and their ability to enter cells in vitro and in vivo compared by Western blotting and immunohistochemistry. The transduction efficiencies and biological activities of the SOD fusion protein with the Tat PTD at either terminus were equivalent and lower than the fusion protein with the Tat PTD at both termini. The availability of a more efficient SOD fusion protein provides a powerful vehicle for therapy in human diseases related to this anti-oxidant enzyme and to reactive oxygen species.

• Journal of Radiation Protection and Research
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• v.30 no.4
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• pp.197-208
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• 2005

Although little information is available on the underlying mechanisms, various genetic factors have been associated with tissue-specific responses to radiation. In the present study, we explored the possibility whether organ specific gene expression is associated with radiosensitivity using samples from brain, lung and spleen. We examined intrinsic expression pattern of 23 genes in the organs by semi-quantitative RT-PCR method using both male and female C57BL/6 mice. Expression of p53 and p21, well known factors for governing sensitivity to radiation or chemotherapeutic agents, was not different among the organ types. Both higher expression of sialyltransferase, delta7-sterol reductase, leptin receptor splice variant form 12.1, and Cu/Zn superoxide dismutase (SOD) and lower expression of alphaB crystalline were specific for spleen tissue. Expression level of glutathione peroxidase and APO-1 cell surface antigen gene in lung tissue was high, while that of Na, K-ATPase alpha-subunit, Cu/ZnSOD, and cyclin G was low. Brain, radioresistant organ, showed higher expressions of Na, K-ATPase-subunit, cyclin G, and nucleolar protein hNop56 and lower expression of delta7-sterol reductase. The result revealed a potential correlation between gene expression patterns and organ sensitivity, and Identified genes which might be responsible for organ sensitivity.

• Journal of Aquaculture
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• v.19 no.3
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• pp.157-165
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• 2006

Expression of major antioxidant enzyme (AOE) including Cu/Zn superoxide dismutase (Cu/Zn-SOD), catalase (CAT), glutathione-S-transferase (GST) and 3 glutathione peroxidase isotypes (GPXs) at mRNA levels during heat stress was examined in mud loach (Misgurnus mizolepis) liver. Based on the semi-quantitative RT-PCR, real-time RT-PCR and/or northern dot blot hybridization, the antioxidant enzyme genes were generally up-regulated during elevation of water temperature from $23^{\circ}C$ up to $32^{\circ}C$. GPXs and SOD displayed the most significant elevation of mRNA levels (up to 3 and 2 folds, respectively) while CAT showed the steady-state expression irrespective of thermal conditions. GST represented the relatively moderate response (1.3-fold increase) in its transcription to thermal stress. The transcriptional activation of AOE genes was not significant at the treatment temperature lower than $29^{\circ}C$. Increased mRNA levels of GPX (extracellular form) and SOD genes in the fish exposed to $32^{\circ}C$ was readily detectable 1 day after exposure to heat stress.

• Journal of Embryo Transfer
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• v.26 no.2
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• pp.135-140
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• 2011

Genistein is a product of naturally occurring isoflavones at relatively high levels in soybeans. The harmful effects of ethanol are attributed to the induction of biological processes which lead to an increase in the generation of reactive oxygen species in fetuses. In this study, we investigated the effects of genistein ($1{\times}10^{-8}$ and $1{\times}10^{-7}\;{\mu}g$/ml) on gene expressions of the representative cellular antioxidative enzymes in ethanol (1 ${\mu}l$/ml)-treated mouse fetuses during the critical period (embryonic days 8.5~10.5) of organogenesis using a semi-quantitative RT-PCR analysis. The mRNA levels of cytosolic glutathione peroxidase (GPx), phospholipid hydroperoxide GPx, cytosolic CU,Zn-superoxide dismutase (SOD), and mitochondrial SOD were significantly decreased in ethanol-treated fetuses. However, the mRNA levels of ethanol plus genistein-treated fetuses were significantly higher than those of ethanol alone fetuses. These results indicate that genistein can up-regulate the expressions of GPx and SOD mRNAs reduced by the ethanol treatment in fetuses.

• 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.

• Journal of Life Science
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• v.11 no.3
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• pp.279-283
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• 2001

Amyotrophic lateral sclerosis(ALS) is a progressive neurologic disorder resulting from the degeneration of upper and lower motor neurons, and is inherited in 10% of cases. About 20% of familial ALS, clinically indistinguishable from sporadic ALS, is caused by mutations of Cu/Zn superoxide dismutase on chromosome 21q22.21 inherited as an autosomal dominant trait. We now report a new locus in the non-SOD1 dominantly inherited ALS. We screened a large ALS family with 11 affected individuals and one obligate gene carrier with genome-wide ABI polymorphic markers using the ABI 377 automated system. No evidence of linkage was obtained with the autosomal markers. We next screened this family with X chromosome markers as there was no evidence of male-to-male tran-smission of the disease. Linkage was established with several X chromosome markers with a lod score up to 3.8; almost the maximum possible score in this family. Our finding imply that a gene for the dominant expression of a neuronal degeneration is coded on X chromosome and raise the question of the role of X-linked genes that escape inactivation in this pathogenesis. More importantly, our finding that a gene causing ALS is localized on X-chromosome has direct investigational relevance to sporadic ALS, where epidemiological studies show male gender predominance(1.3:1) and earlier onset in men by 5-10 years.

• Korean Journal of Community Nutrition
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• v.12 no.4
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• pp.396-404
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• 2007

Increased oxidative stress contributes to the progression of atherosclerosis. We measured serum antioxidant mineral concentrations, capacities of serum antioxidant enzymes and fasting lipid profile in 97 male patients with coronary artery disease (CAD) and 21 male controls. Nutrient intake was assessed by the semi-quantitative food frequency method. CAD patients were divided into single-vessel disease (SVD, n=66) and multi-vessel disease (MVD, n = 31) groups on the coronary angiography. The ratio of serum LDL- to HDL-cholesterol elevated with an increasing number of diseased vessels compared to the control (control < SVD < MVD, p < 0.05). Patients with SVD and MVD had higher levels of serum lipoprotein (a) than the control (p < 0.05). The mean intake of carbohydrate, protein and cholesterol was higher in MVD patients and the intakes of vitamins C and E were lower in MVD and SVD patients than in the control (p < 0.05). Serum copper (Cu) and zinc (Zn) levels were higher in MVD and SVD patients than in the control (Cu: control $75.8{\pm}5.07$, SVD $99.2{\pm}2.90$, MVD $100.1{\pm}2.32{\mu}g/dL$, p<0.01; Zn: $76.8{\pm}5.36$, $119.0{\pm}5.95$, $129.1{\pm}2.70{\mu}g/dL$, p < 0.01). And the ratio of Zn to Cu was higher in SVD and MVD patients than in the control (control $0.78{\pm}0.06$, SVD $0.88{\pm}0.05$, MVD $0.99{\pm}0.04$, P < 0.05). The activity of glutathione peroxidase (GSH-Px) was lower in MVD than in SVD and the control (control $35.13{\pm}1.34$, SVD $35.30{\pm}1.01$, MVD $31.00{\pm}1.04 U/mg$ protein, p < 0.05). The ratio of the activities of superoxide dismutase (SOD) to GSH-Px was higher in MVD than in control and SVD (p < 0.05). In groups with CAD, serum Cu and Zn concentrations and their ratio were changed compared to the control. GSH-Px activity was decreased and the ratio of SOD to GSH-Px was increased in the patients with MVD. The balances between the activities of SOD and GSH-Px should also be considered a risk factor in CAD patients.