• Title/Summary/Keyword: Glutathione S-transferase 유전자

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Nucleotide Sequence and Homology Analysis of phnC Gene Encoding Glutathione S-transferase from Pseudomonas sp.DJ77 (Pseudomonas sp. DJ77에서 Glutathione S-transferase를 암호하는 phnC 유전자의 염기서열과 상동성 분석)

  • 우희종;신명수;김성재;정용제;정안식;박광균;김영창
    • Korean Journal of Microbiology
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    • v.33 no.2
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    • pp.86-91
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    • 1997
  • Pseudomonas sp. DJ77로부터 클로닝된 glutathione S-transferase 유전자(phnC)의 염기서열을 결정하였다. 603bp의 open reading frame(ORF)이 존재하였고 개시코돈 앞에서 Shine-Dalgarno sequence를, 종결코돈 뒤에서는 terminator sequence를 발견하였다. phnC 유전자에서 만들어지는 phnC 단백질은 21,416 Da으로 SDS-polyacrylamide gel 전기영동 결과와 일치하였다. PhnC는 Bulkholderia cepacia LB400, Cycloclasticus oligotrophus RB1의 GST와 각각 53.7%, 49%의 높은 상동성을 나타냈다. 아미노산 서열의 상동성과 필수잔기들의 존재유무로 판단할 때 PhnC GST는 theta class GSTs와 진화적으로 유연관계가 높았지만 alpha, mu, pi, sigma class GSTs에서 구조적, 기능적으로 중요하다고 알려진 아미노산 잔기들이 PhnC GST에도 보존되어 있었다. 또한, phnC 유전자의 위치가 C. oligotrophus RB1, B. cepacia LB400 등의 GST 유전자 위치와 유사하다는 점에서 PhnC 효소는 난분해성 방향족 탄화수소의 분해에 관여하는 것으로 생각된다.

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Association between the Polymorphism of Glutathione S-transferase Genes and Chronic Myeloid Leukemia in Asian Population: a Meta-analysis (아시아인종에서 만성골수성백혈병과 Glutathione S-transferase 유전자 다형성의 메타분석)

  • Kim, Hee Sung
    • The Journal of the Korea Contents Association
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    • v.17 no.10
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    • pp.289-299
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    • 2017
  • To verify the association between susceptibility to chronic myeloid leukemia (CML) and GSTM1, GSTT1 gene polymorphisms in Asian populations, 9 papers published until July 2017 were cited in a meta-analysis. The null present types of the GSTM1, GSTT1 gene were analyzed individually. The significant association was found between CML and GST polymorphism (GSTM1; OR=1.306, 95% CI=1.091-1.563, p=0.004, GSTT1; OR=1.987, 95% CI=1.438-2.746, p=0.000). In addition, there was association between CML and the null type of the combination GSTM1-GSTT1 polymorphisms (OR=4.191, 95% CI=2.833-6.201, p=0.000). Thus, genetic polymorphisms of the GSTM1, GSTT1 and combination GSTM1-GSTT1 polymorphism in Asian populations may be risk factors for CML.

Inductoin of Radioresistance by Overexpression of Glutathione S-Transferase K1 (hGSTK1) in MCF-7 Cells (MCF-7 세포주에서 Glutathione S-Transferase K1 (hGSTK1) 과발현에 의한 방사선 내성의 유도)

  • Kim, Jae-Chul;Shin, Sei-One
    • Radiation Oncology Journal
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    • v.19 no.4
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    • pp.381-388
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    • 2001
  • Purpose : This study was conducted to assess the effects of x-irradiation on the expression of the novel glutathione S-transferase K1 gene. Materials and methods : Human glutathione S-transferase K1 (hGSTK1) DNA was purified and ligated to a pcDNA3.1/Myc-His(+) vector for the overexpression of hGSTK1 gene. MCF-7 cells were transfected with or without the recombinant hGSTK1 gene, and irradiated with 6 MV x-ray. After incubation of 14 days, cell survival was measured and compared. The expression of hGSTK1 and the effect of x-irradiation on hGSTK1 expression were also estimated in MCF-7 cells transfected with or without the hGSTK1 gene by RT-PCR. Results : Following 2 to 12 Gy of x-irradiation, the cell survivals were higher in the MCF-7 cells transfected with the hGSTK1 gene than in those without transfection. Despite the higher cell survival in the hGSTK1-transfected cells, RT-PCR for hGSTK1 mRNA revealed no significant differences according to radiation dose, fractionation, and time after irradiation. Conclusion : The MCF-7 cells transfected with the hGSTK1 gene showed higher cell survival than those without transfection of the gene. The hGSTK1 gene might be associated with the radiosensitivity of MCF-7 cell line and further analysis should be needed.

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Effect of Cadmium on the Expression of ABC Transporters and Glutathione S-transferase in the Marine Ciliate Euplotes crassus (카드뮴이 해양 섬모충(Euplotes crassus)의 ABC Transporters와 GST 유전자 발현에 미치는 영향에 관한 연구)

  • Kim, Hokyun;Kim, Se-Hun;Kim, Ji-Soo;Lee, Young-Mi
    • Journal of Marine Life Science
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    • v.1 no.2
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    • pp.79-87
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    • 2016
  • Heavy metals such as cadmium (Cd) are highly toxic to aquatic organisms and human, even at trace concentration. Herein we investigated the effect of Cd on the gene expression of ATP-binding cassette (ABC) transporters and glutathione S-transferase (GST) in marine ciliate Euplotes crassus. Seven ABC transporters and one GST genes were partially cloned and sequences, and thereafter, transcriptional modulation of these genes after exposure to Cd for 8 h was investigated using quantitative real time RT- PCR (qRT-PCR). As results, sequence analysis and phylogenetic study revealed that E. crassus ABCs are likely typical ABC transports, in particular, B/C family, and GST gene may be similar to GST theta isoform. A significant increase in the expression of ABCs, except for ABCB21 was observed in a concentration dependent manner after exposure to Cd (0.1 and 0.5 mg/l) for 8 h. The GST mRNA level was the highest at 0.5 mg/l Cd and then reduced until control level. These findings suggest that ABCs and GST may be involved in a protective mechanism against Cd-mediated toxicity in E. crassus.

Isolation of Gglutatihone S-Ttransferase(ClGST) Gene from Codonopsis lanceolata (더덕에서 Glutathione S-transferase (ClGST) 유전자의 분리)

  • Kim Jin-Ju;Yang Deok-Chun
    • Korean Journal of Plant Resources
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    • v.18 no.2
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    • pp.240-245
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    • 2005
  • A cDNA clone homologous to glutathione S-transferase gene was isolated and characterized from Codonopsis lanceolata(ClGST). The ClGST is 761 nucleotides long and has an open reading frame of 522 bp with a deduced amino acid sequence of 173 residues. The ClGST shows meaning homology to A. thaliana(AAC63629) $71\%$, C. chinense(CAI51314) $73\%$, E. esula(AAE65767) $75\%$, H. muticus(CAA55039) $70\%$, N. plumbaginifolia(CAA96431) $77\%$, S. commersonii(AAB65163).

Expression of Human p53 Gene as Glutathione S-transferase Fusion Proteins in Escherichia coli (사람의 p53 유전자와 Glutathione S-Transferase와의 융합 단백질의 대장균에서의 발현)

  • 오상진
    • Korean Journal of Microbiology
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    • v.31 no.4
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    • pp.279-285
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    • 1993
  • Alterations of the p53 gene arc among the most frequent genetic changes in human cancer and often result in increased levels of p53 protein within the malignant cells. Detection of accumulated p53 protein can be a useful prognostic tool in human cancer. In order to make polyclonal antibodies for immunohistochemical screening. human p53 gene was expressed in E. coli in the form of GST (glutathione S-transfi.:rase) fusion proteins. Two p53 gene fragments. which were N('()I small fragment encoding amino acid residues of 1-151-: and Ncol large fragment of 159-393. were subeloned into the unique BamHI site present within the pGEX-2T vector using BamHI linker and recombinant plasmids pGTNS and pGTNL were constructed. respectively. The p53 cDNA fragment (from pC53-$SN_3$,) encoding amino acid 38-145 (proline at residue 72) was amplified by polymerase chain reaction(PCR). The amplified DNA was digested with BamHI and Prull and inserted into the BamHI-Smal sites of pG EX-2T and recombinant plasmid pGTBP was constructed. After IPTG induction of these plasmids for 4 hours. fusion proteins were purified from E. coli extracts with glutathione Sepharose beads. The bound proteins were resolved by 10% SDS-polyacrylamide gel electrophoresis and the molecular weights were 54 kDa. 53 kDa and 40 kDa. respectively. Approximately one milligram of fusion proteins were purified from 1 -liter cultures.

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Effect of Bisphenol A on Early Embryonic Development and the Expression of Glutathione S-transferase (GST) in the Sea Urchin (Hemicentrotus pulcherrimus) (말똥성게(Hemicentrotus pulcherrimus)의 초기배아 발생과 glutathione S-transferase (GST)의 발현에 대한 bisphenol A의 영향)

  • Hwang, Un-Ki;Kim, Dae-Han;Ryu, Hyang-Mi;Lee, Ju-Wook;Park, Seung-Yoon;Kang, Han Seung
    • Korean Journal of Environmental Biology
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    • v.32 no.3
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    • pp.234-242
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    • 2014
  • In this study, gametotoxicity and embryotoxicity experiments using Hemicentrotus pulcherrimus were carried out to investigate the ecotoxicological effects of bisphenol A (BPA). We examined the effects of BPA on fertilization and normal embryogenesis at various concentrations (0, 300, 500, 800, 1000, and 1500 ppb). The results demonstrated that the fertilization rates were not changed. The normal embryogenesis rates were gradually decreased in a dose-dependent manner, and were significantly lowered following 800 ppb BPA treatment ($EC_{50}$=1056.1 ppb, 95% Cl=981.8~1163.9 ppb). The observed effective concentration and the lowest observed effective concentration of the normal embryogenesis rate were 500 ppb and 800 ppb, respectively. The embryos showed retarded development at each tested concentration, indicating the fact the embryonic development was delayed due to the increasing concentrations of BPA. Furthermore, we examined the expression of glutathione S-transferase (GST) mRNA at various concentrations of BPA in H. pulcherrimus. Interestingly, it was found that the expression level of GST mRNA was significantly increased in the experimental group exposed to BPA. Based on these results, we suggested that BPA at greater than 800 ppb has a toxic effect during the early embryonic stages of H. pulcherrimus, and GST mRNA may be used as a biomarker for risk assessment of BPA contamination.

Effect of Glutathione S-Transferase Polymorphisms on the Antioxidant System (Glutathione S-Transferase 유전적 다형성이 항산화 체계에 미치는 영향)

  • Jeon, Gyeong-Im;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.6
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    • pp.708-719
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    • 2007
  • Glutathione S-transferase genotypes GSTT1, GSTM1 and GSTP1 were characterized in 104 healthy male and female subjects and compared with parameters of oxidative stress at the level of DNA and lipids, with antioxidant enzymes, and with plasma antioxidants in smokers and non.smokers. Of the 104 subjects studied, 57.4% were GSTT1 present and 47.6% were GSTM1 present. The GSTP1 polymorphisms a and b were represented as follows: a/a, 75.5%; a/b, 21.6%; b/b type, 2.9%. The GSTT1 null genotype was associated with decreased glutathione in erythrocytes and elevated lymphocytes DNA damage. GST-Px was higher in GSTT1 null compared with GSTT1 present type. The homozygous GSTP1 genotype was not associated with any antioxidant status or DNA damage. The difference in plasma ${\alpha}$-carotene and erythrocytes GSH-Px and GST activities between smokers and non-smokers was detected in the GSTT1 null genotype. Plasma ${\gamma}$-tocopherol and ${\beta}$-carotene decreased significantly in smokers having GSTM1 null genotype. When GSTT1 and GSTM1 were combined, plasma lycopene and erythrocyte GST were reduced in smokers in both null types of these genes. As for GSTP1 genotype, plasma ${\alpha}$-carotene and erythrocytes GSH-Px decreased significantly in smokers with GSTP1 b/b, while erythrocytes GSH-Px activities decreased in smokers with GSTP1 a/b. The different ${\beta}$-carotene level between smokers and non-smokers was seen with both GSTP1 a/a and a/b genotype. It seems that polymorphisms in the phase II metabolizing enzyme glutathione S-transferase may be important determinants of commonly measured biomarkers.

Increase of Salt and Low Temperature Tolerance by Overexpressing Glutathione S-Transferase (GST) Gene (염분과 저온에 대한 내성증진을 위한 GST 유전자의 과발현)

  • Jun Chol Kim;Il Seop Kim;Won Hee Kang
    • Journal of Bio-Environment Control
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    • v.11 no.3
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    • pp.139-143
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    • 2002
  • Cotton Glutathione S-Transferase (GST: EC 2.5.1.18) was cloned and overexpressed in tobacco (Nicotiana tabacum) plants. Northern blot analysis confirmed the successful transformation of cotton gst gene in tobacco plant. Type I and Type ll transcript patterns were identified in transgenic tobacco plants and only Type I transcripts were discussed in this paper, The activity of GST in the type II transgenic plants was about 1.5-fold higher than those of the wild type and non-expresser by using 1-chloro-2,4-dinitrobenzene (CDNB) and reduced glutathione as the substrate. The expression of cotton GST in tobacco plants proved that Gh-5 could be translated into functional protein. Type II transgenic plants produced functional GST in the cells. The effects of cotton GST in the seedlings was evaluated by growing the control and transgenic seedlings at $15^{\circ}C$ in the growth chamber in the light. Overexpressors were grown well compared to the control plants (non-expressors). lo test far tolerance to salinity, seeds of Gh-5 overexpressors and the wild type Xanthi seedlings were grown at 0, 50, 100, 150, and 200 mM NaCl solution. Gh-5 transgenic seedlings showed higher growth rate over control seedlings on 50 and 100 mM NaCl solution. There was no difference in growth rate at 150 and 200mM NaCl concentration.

Association between Antipsychotic-Induced Restless Legs Syndrome and Glutathione S-Transferase Gst-M1, Gst-T1 and Gst-P1 Gene Polymorphisms (Glutathione S-Transferase (GST) 유전자 다형성과 항정신병약물로 유발된 하지불안증후군의 연관 연구)

  • Kang, Seung-Gul;Park, Young-Min;Kim, Leen;Lee, Heon-Jeong
    • Sleep Medicine and Psychophysiology
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    • v.22 no.1
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    • pp.25-29
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    • 2015
  • Objectives: The pathophysiology of restless legs syndrome (RLS) has not been fully elucidated. Oxidative stress might play a role in the development of RLS and other antipsychotic-induced side effects such as tardive dyskinesia. In the present study, we investigated whether the glutathione S-transferase (GST) gene polymorphisms are associated with antipsychotic-induced RLS in schizophrenia. Methods: We assessed antipsychotic-induced RLS symptoms in 190 Korean schizophrenic patients using the diagnostic criteria of the International Restless Legs Syndrome Study Group. The GST-M1, GST-T1 and GST-P1 loci were analyzed using PCR-based methods. Results: We divided the subjects into 2 groups: those with RLS symptoms (n = 96) and those without RLS symptoms (n = 94). There were no significant differences in the distributions of the GST-M1 genotypes (${\chi}^2=3.56$, p = 0.059), GST-T1 (${\chi}^2=0.51$, p = 0.476) and GST-P1 (${\chi}^2=0.57$, p = 0.821) between the 2 groups. Comparison of the RLS score among genotypes of the GST-M1 (t = -1.54, p = 0.125), GST-T1 (t = -0.02, p = 0.985) and GST-P1 (F = 0.58, p = 0.560) revealed no significant difference. Conclusion: These data suggest that GST gene polymorphisms do not confer increased susceptibility to RLS symptoms in schizophrenic patients. Future studies are necessary to evaluate the possible influences of other candidate genes involved in the reactive oxygen species system.