• 원예과학기술지
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• 제29권6호
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• pp.623-632
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• 2011

본 연구는 배추에서 항암물질 PEITC의 함량을 높이기 위하여 PEITC 대사과정에서 관련 유전자인 myrosinase (MYR)와 Glutathione S-transferase(GST) 유전자를 분리하고 Agrobacterium tumefacien 형질전환 방법을 통하여 유전자 발현을 조절하였다. 분리된 MYR과 GST의 cDNA는 각각 1647bp와 624bp임을 확인하였고 pET system으로 단백질의 발현을 확인하였다. 형질전환을 위해서 MYR-과발현 벡터와 GST-발현억제 벡터를 제작하였으며 이를 이용하여 배추에 형질전환한 후 PCR 검정을 통해 MYR-과발현 벡터로 형질전환된 개체(IMS) 13개체를 GST-발현억제 벡터로 형질전환된 개체(IGA) 5개체를 선발하였다. 선발된 $T_0$ 개체는 $T_1$ 세대로 진전시켰으며 $T_1$ 형질전환 계통의 서던분석 결과 배추 genome내로 1-4 copy의 T-DNA가 삽입된 것을 확인하였다. 유전자 발현양을 real-time RT PCR로 조사한 결과 IMS는 발현량이 1.03-4.25배 증가하였고 IGA는 26.42-42.22배 감소하였다. IMS와 IGA의 각 계통에서 PEITC의 농도를 GC-MS 방법을 이용하여 확인한 결과 IMS는 PEITC 함량이 형질전환이 되지 않은 대조군에 비해 최대 4.86배까지 증가한 계통을 확인하였고 IGA는 최대 3.89배까지 증가된 계통을 확인하였다. 최종적으로 본 연구를 통하여 항암물질 PEITC량의 증가를 보인 형질전환계통 IMS 1, 3, 5, 12, 15 및 IGA 1, 2, 4를 선발하였다.

• Animal cells and systems
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• 제12권3호
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• pp.149-155
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• 2008

We have isolated and characterized Agrius convolvuli cDNA encoding a c-type lysozyme. The cDNA sequence encodes a processed protein of 139 amino acid residues with 19 amino acid residues amino-terminal signal sequence and 120 amino acid residues mature sequence. The amino acid residues responsible for the catalytic activity and the binding of the substrate are conserved. Agrius lysozyme has a high identity to Manduca sexta. Recombinant A. convolvuli lysozyme was expressed in Escherichia coli BL21(DE3) pLysS cells for pGEX 4T-1 expression vector. Their optimal conditions for the fusion protein expression and purification were screened. Lysozyme gene amplified with primers ACLyz BamHI and ACLyz XhoI was ligated into the pGEX 4T-1 vector, which contained the glutathione S-transferase(GST) gene for fusion partner. The fusion protein was induced by IPTG and identified by SDS-PAGE analysis. Molecular weight of the fusion protein was estimated to be about 45 kDa. Recombinant lysozyme, fused to GST, was purified by glutathion-Sepharose 4B affinity chromatography. Western blot analysis of this protein revealed an immunoreactivity with the anti-Agrius lysozyme.

• Asian Pacific Journal of Cancer Prevention
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• 제15권5호
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• pp.2075-2081
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• 2014

The association between glutathione-S-transferase polymorphisms (GSTM1, GSTT1 and GSTP1) and risk of acute leukemia in Asians remains controversial. This study was therefore designed to evaluate the precise association in 23 studies identified by a search of PubMed and several other databases, up to December 2013. Using random or fixed effects models odds ratios (ORs) with corresponding 95% confidence intervals (CIs) were calculated. Heterogeneity across studies was assessed, and funnel plots were constructed to test for publication bias. The meta-analysis showed positive associations between GST polymorphisms (GSTM1 and GSTT1 but not GSTP1) and acute leukemia risk [(OR=1.47, 95% CI 1.18-1.83); (OR=1.32, 95% CI 1.07-1.62); (OR=1.01, 95% CI 0.84-1.23), respectively] and heterogeneity between the studies. The results suggested that the GSTM1 null genotype and GSTT1null genotype, but not the GSTP1 polymorphism, might be a potential risk factors for acute leukemia. Further well-designed studies are needed to confirm our findings.

• Asian Pacific Journal of Cancer Prevention
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• 제13권10호
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• pp.5037-5042
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• 2012

Background: Tobacco contains agents which generate various potent DNA adducts that can cause gene mutations. Production of DNA adducts may be neutralized by glutathione S transferase (GST) along with other phase I and phase II enzyme systems. The existence of null type of GST among the population increases the susceptibility to various disorders and diseases. The present study focuses on the impact of high tobacco usage and possible null type mutation in GST loci. Methods: Genotypes of GST were detected by multiplex polymerase chain reaction in unrelated 504 volunteers of high tobacco using natives of Gujarat. Allelic frequencies were calculated using Statistical Package for Social Studies-16 software. Hardy Weinberg Equilibrium (HWE) was calculated using Chi square test. Two sided Fisher's significance test was used to compare allelic frequencies of different populations. Results: The frequency of homozygous null genotype of GSTM1 and GSTT1 were 20% (95% CI 16.7-23.9) and 35.5% (95% CI 31.4-39.9) respectively. The GSTM1 and GSTT1 null allele frequency distribution in the Gujarat population was significantly deviating from HWE. GSTT1 null frequency of Gujaratians was significantly higher and different to all reported low tobacco using Indian ethnics, while GSTM1 was not differing significantly. Conclusion: Tobacco usage significantly influences the rate of mutation and frequency of GSTT1 and M1 null types among the habituates. The rate of mutation in GSTT1 loci was an undeviating response to the dose of tobacco usage among the population. This mutational impact of tobacco on GSTT1 postulates the possible gene - environment interaction and selection of null genotype among the subjects to prone them under susceptible status for various cancers and even worst to cure the population with GSTT1 dependent drugs.

• 약학회지
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• 제44권4호
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• pp.300-307
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• 2000

Expression of xenobiotic-metabolizing enzymes can be altered by xenobiotics, which represents changes in the production of reactive metabolic intermediates as well as toxicities in tissues. Metabolic intermediates derived from xenobiotics are considered to produce the reactive oxygen species including drug free radicals and hydroxyl free radicals, which would be ultimately responsible for drug-induced toxicities. The effects of 1,2-benzothiazine anti-inflammatory agents on the expression of xenobiotic-metabolizing enzymes including major cytochrome P450s, microsomal epoxide hydrolase (mEH) and glutathione S-transferase (GST) were studied in the liver with the aim of providing the part of information on potential production of reactive metabolites and hepatotoxicity by the agents. The synthetic compounds 24, 36 and 39 exhibited anti-inflammatory effects in rats as assessed by the Randall-Selitto method. The anti-inflammatory effect was detected as early as at 30 min after gavaging the agents with the ED5O being noted at 80 mg/kg, which was comparable to that of ibuprofen. Treatment of rats with each compound (100 mg/kg, 3d) resulted in no significant induction in the immunochemically-detectable cytochromes P45O 1A1/2, P450 2B1/2, P45O 2 Cl1 and P45O 2El. Changes in the mEN expression were also minimal, as evidenced by both Western blot and Northern blot analyses. Hepatic GST expression was slightly increased by the agents: GST Ya protein and mRNA expression was ～1.5-fold increased after treatment with compounds 24 and 39, whereas GST Yb1/2 and Yc1/2 mRNA levels were elevated 2- to 3-fold. In summary the effects of the synthetic 1,2-benzothiazines on the expression of major P45O, mEH and G57 were not significant, providing evidence that metabolic activation of the agents, potential drug interaction and hepatotoxicity would be minimal.

• Fisheries and Aquatic Sciences
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• 제4권2호
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• pp.88-92
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• 2001

To examine effects of cadmium on the respiratory burst of kidney phagocytes and antioxidant defense in liver, juvenile red seabream Pagrus major were fed a cadmium-incorporated diet $(1g\;CdC1_2/kg\;diet)$. The respiratory burst activity measured by chemiluminescence (CL) was significantly reduced by oral intake of cadmium. Lipid peroxidation in liver expressed as thiobarbituric acid reactive substances (TBARS) was significantly higher in the fish fed a cadmium-incorporated diet than that of the fish fed a control diet both on Day 3 and Day 9. Liver Glutathione S-transferase (GST) activitiy was significantly increased both on Day 3 and Day 9 by feeding a cadmium-incorporated diet, when compared with the controls. From the present results, it can be concluded that oral intake of cadmium in red seabream is associated with marked reduction of respiratory burst capacity of kidney phagocytes which can elevate susceptibility of fish against infecting pathogens. Cadmium administration also elicits significant increment of lipid peroxidation in liver, and fish try to detoxify cadmium by increasing GST activity.

• Toxicological Research
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• 제8권1호
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• pp.131-137
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• 1992

Sprague-Dawley rats aged six weeks divided into four groups and group 1, 2, 3 and 4 of rats were given an intrapertioneal injection of diethylnitrosamine at 200 mg/kg body weight. Group 4 was Control. Two weeks after beginning of the experiment, group 1 of rats were begun to feed on water containing 0.05% phenobarbital sodium as a promoter for six weeks, and CP-2 were intrapertioneally given to rats of group 2(20mg/kg) and group 3(1mg/kg). Three weeks after beginning of the experiment, partial hepatectomy was performed in all rats. Preneoplastic foci were identified histopathologically by glutathione S-transferase placental form (GST-P) activity. In the Immunohistochemical quantitative analysis of carcinogen-induced foci, it was concluded that CP-2 was not carcinogen.

• The Plant Pathology Journal
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• 제29권1호
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• pp.110-115
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• 2013

Theobroxide, a novel compound isolated from a fungus Lasiodiplodia theobromae, stimulates potato tuber formation and induces flowering of morning glory by initiating the jasmonic acid synthesis pathway. To elucidate the effect of theobroxide on pathogen resistance in plants, Nicotiana benthamiana plants treated with theobroxide were immediately infiltrated with Pseudomonas syringae pv. tabaci. Exogenous application of theobroxide inhibited development of lesion symptoms, and growth of the bacterial cells was significantly retarded. Semiquantitative RT-PCRs using the primers of 18 defense-related genes were performed to investigate the molecular mechanisms of resistance. Among the genes, the theobroxide treatment increased the expression of patho-genesis-related protein 1a (PR1a), pathogenesis-related protein 1b (PR1b), glutathione S-transferase (GST), allen oxide cyclase (AOC), and lipoxyganase (LOX). All these data strongly indicate that theobroxide treatment inhibits disease development by faster induction of defense responses, which can be possible by the induction of defense-related genes including PR1a, PR1b, and GST triggered by the elevated jasmonic acid.

• 한국수산과학회지
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• 제36권6호
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• pp.573-577
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• 2003

Effects of 4-nonylphenol (4-NP) on mixed function oxygenase (MFO) in the kidneies of rockfish (Sebastes schlegeli) were investigated. The cytochrome P45O (CYP) contents and NADPH cytochrome P450 reductase, NADH cytochrome b5 reductase and 7-ethoxyredorufin-O-deethylase (EROD) activities of microsome, glutathione S-transferase (GST) activity of cytosol in rockfish exposed to 4-NP for 7 days using an intraperitoneal injection (25 mg/kg) were quantitatively determined. The GST activity of rockfish exposed to 4-NP were higher, up to 5.2 times higher, than those in the control fish. The activities of NADPH-and NADH-dependent reductases were inhibited. On the other hand, CYP contents and EROD activity of the 4-NP exposed fish demonstrated neither an increasing or decreasing trend.

• 대한의생명과학회지
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• 제26권3호
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• pp.201-209
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• 2020

The effect of mulberry leaves and yacon tuber extracts (MYE) on antioxidant was tested in this study. The present study investigated the in vivo effects of the anti-oxidative effect of MYE on catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase (GST), glutathione peroxidase (GSH-Px), and thiobarbituric acid reactive substances (TBARS). The seven-day acclimation of the mice was divided into six groups: Normal diet group (NOR), high fat diet group (HFD), high fat diet with 0.5% hydroxycitric acid group diet group for positive group (HHCA), high fat diet with 1% mulberry leaf and 1% yacon diet group (MYE-1), high fat diet with 3% mulberry leaf and 3% yacon group (MYE-3) and high fat diet with 5% mulberry leaf and 5% yacon group (MYE-5). The effect of serum antioxidant in the catalase of MYE-1, MYE-3, and HHCA comparing to HFD by 31.0%, 27.7% and 45.2%, respectively (P<0.05~0.01). The effect on hepatic antioxidant in the catalase of HFD was significantly increased 3.7 (77.3%) times than that of NOR (P<0.01). But, the activities of catalase were decreased significantly in MYE-1, MYE-3, MYE-5 and HHCA by 21.7%, 24.2%, 24.9%, and 28.8% compared to HFD, respectively. GSH-Px was significantly decreased in MYE-1, MYE-3, MYE-5 and HHCA by 15.5%, 37.1%, 23.4%, and 23.7% compared to HFD, respectively (P<0.05). The activities of CAT, SOD, GST, GSH-Px, and TBARS were more significantly decreased in MYE-1 and MYE-3 than those of HFD and HHCA. MYE have shown significant effects on anti-oxidative function against high fat diet.