• Animal cells and systems
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• v.16 no.6
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• pp.455-461
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• 2012

Amongst the various antimicrobial peptides, lysozyme plays a central role in initiating and maintaining the antibacterial defense response of insect. Here we propose the biosynthesis and refolding of recombinant lysozyme in Escherichia coli expressed in inclusion body form. The Agrius lysozyme gene was amplified using gene specific primers and then ligated into the pGEX-4T-1 vector, which contained the glutathione S-transferase (GST) gene as a fusion partner. A recombinant lysozyme was expressed in E. coli Rosetta cells using a pGEX-4T-1 expression vector, and the fusion protein was induced by ioporpyl-${\beta}$-D-thiogalactopyranoside (IPTG). The recombinant protein produced as an inclusion body was resolubilized in solubilization buffer, and the resultant solution was dialyzed in refolding buffer. After thrombin cleavage, the recombinant lysozyme was purified by ion exchange chromatography and reverse phase chromatography. The recombinant lysozyme was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and immunoreactivity against the anti-Agrius lysozyme was observed by western blot analysis of this protein. The recombinant lysozyme displayed antibacterial activity against Bacillus megaterium and Micrococcus luteus, which was confirmed by the inhibition zone assay.

• Journal of Plant Biology
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• v.38 no.1
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• pp.39-45
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• 1995

Glycine soja Sieb. et Zucc., a predominantly selfing annual, has been served as a reservoir of germplasm for soybean, G. max (L.) Merr., cultivar improvement. This study describes the levels and distribution of genetic variation within and among 22 Korean populations of G. soja using starch gel electrophoresis. The species maintains very similar levels of genetic variability within populations observed in most other annuals. At the population level, the mean percent of polymorphic loci (P) was 32.6%, mean number of allele per locus (A) was 1.32, and mean expected heterozygosity (He) was 0.112. In addition, total genetic diversity (HT) calculated only for polymorphic loci was 0.347. However, significant differences in allele frequencies among populations were found for all loci (P<0.001 in each case) and, on average, about 70% of the total variation in the species is common to all populations. Indirects estimate of the number of migrants per generation (Nm=0.58, calculated from mean GST) indicates that gene flow is low among Korean populations of the species. In addition, analysis of fixation indices revealed a substantial heterozygote deficiency in most populations and at all loci. This indicates that most populations sampled may have been substructed largely due to inbreeding (predominantly selfing) and restricted gene flow, coupled with founder effect and genetic drift. Considering a high genetic divergence among populations, it is recommended that several Korean populations of the species should be preserved, especially such as populations in the eastern and southeastern Korean peninsula with high variation.

• Bulletin of the Korean Chemical Society
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• v.28 no.4
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• pp.574-580
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• 2007

Full-length chloride intracellular channel protein 1 (CLIC1) is a member of the family of proteins related to bovine chloride intracellular channel p64. Mutations in the SEDL gene cause spondyloepiphyseal dysplasia tarda (SEDT), a rare X-linked chondrodysplasia. The link between the intracellular chloride channels and SEDL is an important step toward understanding their functional interplay. In the present study, CLIC1 protein was subcloned into the pGEX-KG vector and overexpressed in XL-1 blue cells. We developed a large-scale expression system composed of glutathione S-transferase (GST) fused with a 240-amino-acid CLIC1 protein in Escherichia coli. The soluble CLIC1 protein was successfully purified to homogeneity, and its purity, identity, activity and conformation were determined using SDS-PAGE, MALDI-MS, biophotometer and circular dichroism spectroscopic studies. The binding of both CLIC1 and SEDL proteins in vitro was detected by BIAcore biosensor and fluorescence measurements.

• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.1
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• pp.205-211
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• 1999

Effects of Houttuynia cordata ethanol extracts on lipid metabolism and antioxidant enzymes in Sprague Dawley male rats were investigated. High fat used in the diet mixture included 10% of lard, 1% of cholesterol and 0.25% of sodium cholate. Total serum cholesterol contents of the rats fed Houttuynia cordata extracts were decreased compared to the control. On the other hand, HDL cholesterol contents were increased along with the decrease of athrogenic index. When high fat diet was fed, total serum cholesterol contents were significantly increased(p<0.01) with the athrogenic index increase of four times of the control. With the administration of Houttuynia cordata extract HDL cholesterol was increased by 53% in the high fat diet group. Antioxidant enzymes including GST and catalase activities were increased comparing the control. On the otherhand, the extracts lowered phospholipid(p<0.01), GOT, GPT, Cu,Zn SOD and alkaline phosphatase enzyme activities in the serum which are related to the liver functions. Therefore, the above results suggest that Houttuynia cordata ethanol extracts can help to maintain normal liver functions and help to protect from peroxidative damages caused by excess dietary fat intake.

• Journal of Fisheries and Marine Sciences Education
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• v.28 no.4
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• pp.903-912
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• 2016

Hagfish which belongs to the chordate contact cyclostomata, is important phylogenetic relationship between vertebrate and invertebrate. Cathepsins of the cysteine protease family have traditionally been thought to play a major role in intracellular protein degradation and turnover in lysosomes. In this study, Catepsin L was cloned from Inshore hagfish (Eptatretus burgeri), the cDNA encoding ORF of the Eptatretus burgeri Cathepsin L (EbCtL) is 978 bp. The cDNA encoding proEbCtL was expressed in Escherichia coli strain BL21(DE3) using the pGEX-4T-1 expression vector system. The recombinant proEbCtL protein was overexpressed as a approximately 55 kDa fusion protein. The overproduced soluble GST-fusion protein was then applied to glutathione-Sepharose 4B column chromatography; the sample harboring the fusion protein evidenced a high degree of purity when analyzed via SDS-PAGE and Western blot analysis. Its activity was quantied by cleaving the synthetic peptide Z-FR-AMC, Z-LLE-AMC, and Suc-AAF-AMC, and the optimal pH for the protease activity was 8, 9.5, and 9, respectively.

• Biomolecules & Therapeutics
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• v.10 no.4
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• pp.287-292
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• 2002

To find new inhibitory effects from oriental drugs, Albizziae root was extracted in methanol and the extracted was stepwisely fractionated by hexane, chloroform, ethylacetate, butanol and water. In cytotoxic effect of Albizziae root fractions against cancer cell lines including human hepatoma cells(HepG2) were investigated. Expecially the butanol fraction exhibited a inhibition effects on the growth of human hepatoma cells(HepG2). It inhibited of HepG2 cells with the value of IC50. The activities of qutathione after B(a)P treatment were markedly decreased than control, but those levels were increased by the treatment of Albizziae root methanol fraction. The activity of glutathione-S-transferase after B(a)P treatment were markedly decreased than control, but those levels were increased by the treatment of Albizziae root methanol traction. Induction of phase II enzymes is a major mechanism of chemoprevention. The induction levels of quinone reductase(QR) activity in cultured murine hepatoma(Hepa IcIc7)cell by methanol extract of Albizziae root were measured. Among the tested tractions, the extracts of butanol were found to induce QR activities over 2.8 fold than control. These results suggest that Albizziae root has chemopreventive Potential by inducing QR activities and GST levels and increasing GSH

• Journal of the East Asian Society of Dietary Life
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• v.20 no.1
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• pp.37-45
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• 2010

The effects of an Acanthopanacis cortex water extract on lipid levels, lipid peroxide, total antioxidant status and antioxidant enzyme activities were evaluated in rats fed one of the following diets for six weeks: normal diet and deionized water (ND), normal diet and Acanthopanacis cortex water extract (NDC), high fat diet and deionized water (HFD), high fat diet and Acanthopanacis cortex water extract (HFDC). The food intakes were significantly lower, but the food efficiency ratios were significantly higher in the high fat diet groups. The level of HDL-cholesterol in the plasma was significantly increased and the levels of LDL-cholesterol and triglyceride in the plasma were significantly decreased by the Acanthopanacis cortex water extract in the high fat diet groups. As a a result, the AI (atherogenic index) and CRF (cardiac risk factor) were significantly lower in the high fat diet groups that were treated with Acanthopanacis cortex water extract. The triglyceride and the total cholesterol of the liver were also significantly upregulated in the high fat diet groups, while the total cholesterol of the liver decreased in response to treatment with Acanthopanacis cortex water extract (HFDC). The plasma and liver concentrations of thiobarbituric acid reactive substances (TBARS) were significantly reduced by the addition of Acanthopanacis cortex water extract to the normal diet groups. The total antioxidant status (TAS) in the plasma was significantly upregulated by adding Acanthopanacis cortex water extract to the high fat diet groups. The activities of SOD, catalase and GST were also significantly higher in the Acanthopanacis cortex water extract groups when compared to the ionized water groups. The activity of GSH-Px and the concentration of GSH in the liver were significantly higher following the addition of Acanthopanacis cortex water extract to the high fat diet groups. Taken together, these results suggest that a supplementation of the diet of rats fed a high fat diet with Acanthopanacis cortex water extract improves lipid metabolism, reduces lipid peroxide and improves the activities of antioxidant enzymes, which may have favorable effects on antioxidant systems by improving the total antioxidant status (TAS).

• Journal of Food Hygiene and Safety
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• v.8 no.1
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• pp.37-53
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• 1993

F. moniliforme MRC 826, a common fungal contaminant of com, has been known to produce a group of mycotoxins, the fumonisins. By thin layer chromatography, fumonisin $B_{1}$ was detected in the F. moniliforme MRC 826 com culture material(CM) extracts. This study was performed to compare the toxicity and carcinogenicity of F. moniliforme MRC 826 CM with those of aflatoxin $B_1(AFB_1)$ in rats. The toxicity was tested over a period of 7 days in ten female Sprague-Dawley (SD) rats. Treatment group were fed a 1 : 1 mixture(wt/wt) of ground CM and basal diet in powder form, while other negative control group were given basal diet alone. The principal pathological changes in rats treated with 50% CM were hepatocellular hydropic degeneration and renal tubular necrosis. The cancer-promoting activity of CM was evaluated in the rat liver diethylnitrosamine-two thirds partial hepatectomy(DEN-PH) model for carcinogenesis. 70 male SO rats(ca. 170 g) were randomized into 5 groups. Group I served as the positive controls and received the basal diet containing 2 ppm $AFB_{1}$ group 2 received 5% CM, group 3 received 2.5% CM, group 4 received 5% normal com and group 5 received 2.5% normal com. 5% treated group showed cancer promoting activity in rat liver using DEN as initiator and the induction of glutathione S-transferase placental form positive foci as an end point after 6 weeks of promotion.

• Journal of Life Science
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• v.28 no.7
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• pp.765-771
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• 2018

Peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) is a key transcription factor that regulates adipogenesis, and epigenetic control of $PPAR{\gamma}$ is of great interest in obesity-inhibition research. Our previous study showed that CACUL1 (CDK2-associated cullin domain 1) acts as a corepressor that inhibits $PPAR{\gamma}$ transcriptional activity and adipocyte differentiation. Here, we investigated the roles of protein arginine methyltransferase 5 (PRMT5), a novel binding partner of CACUL1, in regulating $PPAR{\gamma}$. The interaction between PRMT5 and CACUL1 was shown by immunoprecipitation assay in vivo and GST pulldown assay in vitro. As shown by luciferase reporter assay, PRMT5 and CACUL1 cooperated to inhibit the transcriptional activity of $PPAR{\gamma}$. The suppressive role of PRMT5 in adipogenesis was examined by Oil Red O staining using 3T3-L1 cells, which stably overexpress or deplete PRMT5. Overexpression of PRMT5 suppresses $PPAR{\gamma}$-mediated adipogenesis, whereas PRMT5 knockdown increases lipid accumulation in 3T3-L1 cells. Consistently, PRMT5 attenuates the expression of Lpl and aP2, the target genes of $PPAR{\gamma}$, as demonstrated by RT-qPCR analysis. Overall, these results suggest that PRMT5 interacts with CACUL1 to impair the transcriptional activity of $PPAR{\gamma}$, leading to the inhibition of adipocyte differentiation. Therefore, the regulation of PRMT5 enzymatic activity may provide a clue to develop an anti-obesity drug.

• Journal of the Korean Society of Food Culture
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• v.17 no.4
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• pp.377-386
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• 2002

The purpose of this study was to investigate the effects of dietary Indongcho(L. japonica Thunb) powder on blood glucose, serum lipid levels and antioxidative enzymes in normal and streptozotocin(STZ)-induced diabetic rats. Four groups of rats(3-week-old inbred Sprague-Dawley male rats) were normal rats fed control diet(NC), diabetic rats fed control diet(DC), normal rats fed Indongcho powder diet(NI), and diabetic rats fed Indongcho powder diet(DI). Diabetes was induced by single injection of streptozotocin(60mg/kg B.W., i.p.). The animals were fed ad libium each of the experimental diet for 5 weeks. Food and water intakes were determined everyday. Blood glucose and serum total cholesterol levels were determined every week. After 5 weeks the animals were sacrificed and activities of antioxidant enzymes and lipid peroxidation products were determined in their liver and kidney homogenates. We also determined serum concentrations of total lipid(TL), total cholesterol(TC), triglycerides(TG) and HDL-cholesterol(HDL-C). Blood sugar and water intake were higher in diabetic group(DC and DI group) than normal group(NC and NI group) and were not significantly decreased by dietary Indongcho intake. Body weight gain and FER(feed efficiency ratio) were reduced by STZ treatment. But, Final body weight was recovered by Indongcho-contained diet. LHR(LDL-cholesterol/HDL-cholesterol) of the DI g re up was significantly lower than the other experimental groups(NC, NI and DC groups). The hepatic glucose 6-phosphatase(G6Pase) activity of the groups fed Indongcho diet(NI and DI group) was lower than the groups fed control diet(NC and DC group) and the G6Pase activity of NI group was recovered to the normal levels(p<0.05). However, The glutathione peroxidase(GPx) and glutathione reductase(GR) activities in liver and G6Pase activity in kidney were not statistically different between the control and diabetic control groups. Renal GST activity of the DI group was recovered by Indongcho intake. In conclusion, these results confirm oxidative stress in the liver and kidney of rats with STZ diabetes and antioxidative effect of Indongcho.