• Journal of Microbiology and Biotechnology
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• v.22 no.6
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• pp.866-871
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• 2012

A novel cholesterol-based cationic lipid containing a tri-2-hydroxyethylamine head group and ether linker (Chol-THEA) was synthesized and examined as a potent gene delivery vehicle. In the preparation of cationic liposome, the addition of DOPE as helper lipid significantly increased the transfection efficiency. To find the optimum transfection efficiency, we screened various weight ratios of DOPE and liposome/DNA (N/P). The best transfection efficiency was found at the Chol-THEA:DOPE weight ratio of 1:1 and N/P weight ratio of 10~15. Most of the plasmid DNA was retarded by this liposome at the optimum N/P weight ratio of 10. The transfection efficiency of Chol-THEA liposome was compared with DOTAP, Lipofectamine, and DMRIE-C using the luciferase assay and GFP expression. Chol-THEA liposome with low toxicity had better or similar potency of gene delivery compared with commercial liposomes in COS-7, Huh-7, and MCF-7 cells. Therefore, Chol-THEA could be a useful non-viral vector for gene delivery.

• Proceedings of the Microbiological Society of Korea Conference
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• 2000.10a
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• pp.15-23
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• 2000

The subcellular localization of the SopB protein, which is encoded by the Escherichia coli F plasmid and is involved in the partition of the single-copy plasmid, was directly visualized through the expression of the protein fused to the jellyfish green fluorescent protein (GFP). The fusion protein was found to localize to positions close but not at the poles of exponentially growing cells. Examination of derivatives of the fusion protein lacking various regions of SopB suggests that the signal for the cellular localization of SopB resides in a region close to its N terminus. Overexpression of SopB led to silencing of genes linked to, but well-separated from, a cluster of SopB-binding sites termed sopC. In this SopB-mediated repression of sopC-linked genes, all but the N-terminal 82 amino acids of SopB can be replaced by the DNA-binding domain of a sequence-specific DNA -binding protein, provided that the sopC locus is also replaced by the recognition sequence of the DNA-binding domain. These results suggest a mechanism of gene silencing: patches of closely packed DNA-binding protein is localized to specific cellular sites; such a patch can capture a DNA carrying the recognition site of the DNA -binding domain and sequestrate genes adjacent to the recognition site through nonspecific binding of DNA.

• Korean Journal of Pharmacognosy
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• v.52 no.2
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• pp.105-111
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• 2021

To investigate the antioxidant activities of Pinus densiflora needle (Pinaceae), which is widely used as a traditional medicine in Korea, Caenorhabditis elegans model system was used. The n-butanol fraction of P. densiflora needle methanol extract showed the most potent DPPH radical scavenging and superoxide quenching activities in a dose-dependent manner. The n-butanol fraction was measured for the activities of superoxide dismutase (SOD), catalase, and oxidative stress tolerance by using C. elegans with reactive oxygen species (ROS) level. In addition, to see that the regulation of the stress response gene is responsible for the increased stress tolerance of C. elegans treated by the butanol extract, SOD-3 expression was measured using a transgenic strain. As a result, the P. densiflora needle n-butanol fraction increased SOD and catalase activities, and decreased ROS accumulation, dose-dependently. Furthermore, the n-butanol fraction-treated CF1553 worm showed higher SOD-3::GFP intensity than the control.

• Korean Journal of Pharmacognosy
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• v.50 no.2
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• pp.96-101
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• 2019

Methanol extract of Helianthus tuberosus L. (Compositae) flower was investigated to research the anti-oxidative activity by using a Caenorhabditis elegans model system. Ethyl acetate soluble fraction of the methanol extract showed the most potent DPPH radical scavenging activity. The ethyl acetate fraction was measured on its activities of superoxide dismutase (SOD), catalase, and oxidative stress tolerance with reactive oxygen species (ROS) level in C. elegans. Furthermore, in order to verify if regulation of stress-response gene is responsible for the increased stress tolerance of C. elegans which treated by the ethyl acetate fraction, we checked SOD-3 expression using a transgenic strain. Consequently, the ethyl acetate fraction of H. tuberosus flower increased the catalase and SOD activities in a dose-dependent manner in C. elegans, reduced ROS accumulation dose-dependently. Besides, the ethyl acetate fraction-treated CF1553 worms showed higher SOD-3::GFP intensity than the control group.

• Korean Journal of Pharmacognosy
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• v.49 no.4
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• pp.322-327
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• 2018

Methanol extract of Salvia miltiorrhiza Bunge (Labiatae) root was investigated to research the anti-oxidative activity, by using a Caenorhabditis elegans model system. The methanol extract of this plant showed significant DPPH radical scavenging and superoxide quenching activities. Ethyl acetate soluble fraction of the methanol extract that showed the most potent DPPH radical scavenging and superoxide quenching activities. The ethyl acetate fraction was tested on its activities of superoxide dismutase (SOD), catalase, and oxidative stress tolerance in C. elegans. Furthermore, in order to see if regulation of stress-response genes is responsible for the increased stress tolerance of the ethyl acetate fraction treated C. elegans, we checked SOD-3 expression using a transgenic strain. Consequently, the ethyl acetate fraction of S. miltiorrhiza root increased the catalase and SOD activities in a dose-dependent manner in C. elegans. Besides, the ethyl acetate fraction-treated CF1553 worms showed higher SOD-3::GFP intensity than the non-treated ones.

• Journal of Plant Biotechnology
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• v.46 no.2
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• pp.88-96
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• 2019

Terpenes constitute a large class of secondary metabolites in plants. The Oryza sativa terpene synthase is a vital gene in plant defense response. In this study, the molecular and physiological functions of Oryza sativa terpene synthase 30 (OsTPS30, LOC_Os08g07080) were investigated after exposure of the seeds and plants to gamma-rays. The OsTPS30 expression was slightly induced at 200 Gray (Gy), but was significantly induced at 400 Gy. The total terpenoid was synthesized more in OsTPS30-overexpressing (OX-OsTPS30) Arabidopsisthaliana plants than in wild-type (WT) plants. The OX-OsTPS30 plants exhibited resistance to gamma-rays, as compared to WT. The OX-OsTPS30 plants had significantly increased height and weight after gamma irradiation. Additionally, the activity of antioxidant enzymes was increased more in OX OsTPS30 plants than in WT plants after gamma irradiation. Furthermore, the OsTPS30-GFP fusion protein was mostly localized in the chloroplast, suggesting that OsTPS30 is putative MEP pathway-related terpene synthase.

• Korean Journal of Pharmacognosy
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• v.53 no.4
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• pp.207-212
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• 2022

Through the Caenorhabditis elegans model system, the antioxidant activity of methanol extract of Ixeis dentata was investigated. The ethyl acetate soluble fraction of the I. dentata methanol extract showed the best DPPH radical scavenging activity. The ethyl acetate fraction was measured for the activity of superoxide dismutase (SOD), catalase, and oxidative stress tolerance by using C. elegans along with reactive oxygen species (ROS) level. In addition, to confirm that the regulation of the stress response gene is responsible for the increased stress tolerance of C. elegans treated by the ethyl acetate fraction, SOD-3 expression was measured using a transgenic strain (CF1553). As a result, the ethyl acetate fraction increased SOD and catalase activity, and decreased ROS accumulation in a dose-dependent manner. In addition, the ethyl acetate fraction-treated CF1553 worm showed higher SOD-3::GFP intensity than the control worm.

• Fisheries and Aquatic Sciences
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• v.25 no.2
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• pp.90-100
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• 2022

Cardiovascular diseases (CVDs) have posed serious public health problems, accounting for nearly 30% of mortality worldwide and their incidence is still increasing. Therefore, new treatment resources are necessary to prevent or manage the ever-increasing population of patients with CVDs. Sea cucumber is well known for its medical and health benefit effects, but it is not well known what/how effect it has on vascular disease. In the present study, we examined the protect effect of sea cucumber, Apostichopus japonicus 80% ethanol extract (AJE) on zebrafish embryo with the stimulation of free fatty acid, palmitate (PA). In vivo study showed that AJE can attenuate PA-induced toxicity through relieving the rapid heartbeat, increasing the survival rate and reducing the malformation in both wild type and Tg (fli1a:eGFP) transgenic zebrafish lines. Additionally, compare with PA treated embryos, the yolk sac area, body length, axial vascular segment (AVS) and intersegmental vessel (ISV) of the co-treatment group of AJE and PA were comparable to the control group. Moreover, AJE lowered the expression of inducible nitric oxide synthase (iNOS), nitric oxide (NO) and inflammation-related genes induced by PA, and inhibited PA-induced vascular development disorders. Our data preliminarily verify that AJE could be a candidate resource for the prevention or therapy of CVDs.

• Korean Journal of Pharmacognosy
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• v.53 no.2
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• pp.96-101
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• 2022

Caenorhabditis elegans model system was used to investigate the antioxidant activity of methanol extract of Melampyrum roseum (Scrophulariaceae). The ethyl acetate soluble fraction of the M. roseum methanol extract showed the best DPPH radical scavenging activity. The ethyl acetate fraction was measured for the activity of superoxide dismutase (SOD), catalase, and oxidative stress tolerance by using C. elegans along with reactive oxygen species (ROS) level. In addition, to confirm that the regulation of the stress response gene is responsible for the increased stress tolerance of C. elegans treated by the ethyl acetate fraction, SOD-3 expression was measured using a transgenic strain. As a result, the ethyl acetate fraction increased SOD and catalase activity, and decreased ROS accumulation in a dose-dependent manner. In addition, the ethyl acetate fraction-treated CF1553 worm showed higher SOD-3::GFP intensity than the control worm.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.51-51
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• 2001

Cocaine and amphetamine-regulated transcript (CART) is a satiety factor that is regulated by leptin. It was reported that the mice intracerebroventricularly injected with CART showed behavioral changes resembled with the typical behavioral alterations found in the mice carrying disorders in the brain serotonergic (5-HT) system. Hence, this study was conducted to find out the relationships between CART and 5-HT. We first examined the mRNA levels of CART after the injections of para-chlorophenylalanine (pCPA, 300 mg/kg i.p., single injection or daily for three consecutive days) in the rat brains by in situ hybridization using the mouse CART cDNA probe cloned in our laboratory. Systemic administrations of pCPA, a potent inhibitor of tryptophan hydroxylase, the rate limiting enzyme of 5-HT biosynthesis, acutely depletes the brain 5-HT transporter (5-HTT) in the dorsal raphe nucleus (DRN), which reuptakes terminal 5-HT. Results indicated that the mRNA level of CART significantly decreased in the arcuate nucleus, paraventricular nucleus, and lateral hypothalamic nucleus by three days of daily injection with pCPA with no noticeable change detected 24 hrs after the single injection. The message levels of 5-HTT in DRN decreased in both single and three days of injections. Secondly, to investigate whether CART affect to 5-HT, mouse genomic CART gene, which is consist of 3 exons and 2 introns and mouse neurofilament light (NF-L) chain promoter were cloned. Then, we constructed neuron specific expression vector, which was transfected into HeLa cell using lipid-mediated transfection system. Expression of GFP and CART linked to NF-L-chain promoter in the transfected HeLa cell were detected by using fluorescent microscope and RT-PCR. These results confirmed normal expression of DNA constructs in vitro. Then, to increase brain specific expression of CART in vivo transgenic mice carrying CART gene controlled the deleted NF-L-chain promoter were generated by the DNA microinjection into pronuclei of fertilized embryos. Transgenic mice were detected by Southern blot. Further study is necessary to examine CART expression and 5-HTT in these transgenic mice. Therefore, these results suggest that there maybe a positive molecular correlation between CART and 5-HT in responding to the stimuli.