• 제목/요약/키워드: S-transferase

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

  • 오상진
    • 미생물학회지
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    • 제31권4호
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    • pp.279-285
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    • 1993
  • p53 유전자의 변화는 인간의 여러 암에서 가장 흔하게 발견되며 종양세포내에서는 이러한 변형된 p53 단백질의 양의 증가가 초래된다. 세포내에 축적된 p53 단백질의 발견은 인간의 암증세를 판단할 유용한 기중이 되기도 한다. 본 연구에서는 이러한 면역조직화학 검사에 쓰일 수 있는 폴리클로날 항체를 만들기 위햐여 사람의 p53 유전자를 glutathione S-transferase 와의 융합 단백질의 형태로서 대장균내에서 발현시켰다. p53 의 아미노산 1-158번을 코딩하고 있는 NeoI fragment 와 아미노산 159-393 번을 코딩하는 NocI-BamHI fragment 를 BamHI linker 를 이용하여 in frame 으로 pGEX-2T 의 BamHI 자리에 삽입하여 재조합 플라스미드 pGTNS 와 pGTNL 을 각각 만들었다. 또 PCR 에 의한 증폭에 의햐여 아미노산 38-145번을 코딩하는 유전자 부위를 증폭하였으며 BamHI 과 PvuII 로 절단하여 pGEX-2T의 BamHI 과 SmaI 자리에 삽입함으로써 pGTBP 를 제조하였다. 이들 재조합 균주들을 IPTG 로 4시간 induction 한 후 세포 추출물로부터 glutathione Sepharose bead 를 이용하여 융합단백질을 분리하였다. Bead 에 결합된 단백질은 10% SDS-polyacrylamide gel 에서 전기영동하였으며, 각각의 분자량은 54 kDa, 53 kDa 와 40 kDa 였다. 이러한 방법으로 1리터 배양으로부터 약 1mg 의 단백질을 정제하였다.

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

  • 강승걸;박영민;김린;이헌정
    • 수면정신생리
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    • 제22권1호
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    • pp.25-29
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    • 2015
  • 목 적 : 하지불안증후군(restless legs syndrome ; RLS)의 병인은 아직 불명확하지만, 유전적 질환으로 알려져 있다. 산화스트레스는 RLS, 지연성운동장애, 파킨슨병, 뚜렛장애 등의 운동장애에서 주요한 원인 중의 하나로 생각되고 있다. 본 연구에서는 조현병환자에서 항정신병약물에 의해 유발된 RLS 증상이 산화손상의 해독효소인 glutathione S-transferase (GST) 유전자의 다형성과 연관이 있는지를 밝히고자 하였다. 방 법 : International Restless Legs Syndrome Study Group의 진단기준으로 190명의 한국인 조현병 환자들을 대상으로 RLS에 대해서 평가하였다. 유전자형분석은 중합효소연쇄반응기법을 사용하여 GST-M1, GST-T1, GST-P1의 세 가지 단일염기다형성(single nucleotide polymorphism, SNP)에 대해서 시행되었다. 결 과 : RLS 증상군 96명과 무증상군 94명으로 피험자들을 분류하였다. GST-M1 (${\chi}^2=3.56$, p = 0.059), GST-T1 (${\chi}^2=0.51$, p = 0.476), GST-P1 (${\chi}^2=0.57$, p = 0.821)의 유전자형 빈도에 두 군간에 통계적으로 유의한 차이가 없었다. 유전자형에 따른 RLS 척도의 점수도 GST-M1 (t = -1.54, p = 0.125), GST-T1 (t = -0.02, p = 0.985), GST-P1 (F = 0.58, p = 0.560)의 세 가지 SNP에서 통계적으로 유의한 차이를 보이지 않았다. 결 론 : 본 연구의 결과 GST 유전자 다형성이 항정신병약물로 유발된 RLS 증상 발생의 민감성을 증가시킨다는 증거는 발견할 수 없었다. 산화손상과 관련된 다른 후보 유전자들에 대한 향후 연구가 필요할 것으로 사료된다.

Overexpression of Cotton Glutathione S-Transferase (GST) cDNA and Increase of low Temperature and Salt Tolerance in Plants

  • Kang, Won-Hee;Jong Hwa kim;Lim, Jung-Dae;Yu, Chang-Yeon
    • Journal of Plant Biotechnology
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    • 제4권3호
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    • pp.117-122
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    • 2002
  • Cotton Glutathione S-Transferase(GST: EC 2.5.1.18) was cloned and Gh-5 cDNA was overexpressed in tobacco (Nicotiana tabacum) plants. The transformation of cotton GST in tobacco plant was confirmed by northern blot analysis. Type I and Type II transcript patterns were identified in Gh-5 transgenic tobacco plants. Type I transcripts was only discussed in this paper. Glutathione and 1-chloro-2,4-dinitrobenzene (CDNB) were used as the substrates, and the activity of GST in the type I transgenic plants was about 2.5-fold higher than the non-expressers and wild type tobacco plants. The expression of cotton GST in tobacco plants proved that Gh-5 could be translated into functional protein. Type I transgenic plants produced functional GST in the cells. Type I showed higher GST specific activity than Type II in the transgenic plants. Control and transgenic seedlings were grown in the growth chamber and under the light at 15$^{\circ}C$, and the effects of cotton GST in the seedlings was evaluated. The growth rate of Gh-5 overexpressors was better than the control and non-transgenic tobacco plants. Salinity tolerance was also analyzed on the seeds of transgenic plants. Seeds of Gh-5 overexpressors and the wild type tobacco seedlings were germinated and grown at 0, 50, 100, 150, and 200 mM NaCl solution. Gh-5 transgenic seedlings showed higher growth rate over control seedlings at both 50 and 100 mM NaCl solution. But at 0, 150, and 200 mM NaCl concentration, the difference in growth rate was not detected.

Lack of Association of Glutathione S-transferase M3 Gene Polymorphism with the Susceptibility of Lung Cancer

  • Feng, Xu;Dong, Chun-Qiang;Shi, Jun-Jie;Zhou, Hua-Fu;He, Wei;Zheng, Bao-Shi
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권9호
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    • pp.4465-4468
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    • 2012
  • Objective: The conclusions of published reports on the relationship between the glutathione S-transferase M3 (GSTM3) A/B gene polymorphism and the risk of lung cancer are still debated. This meta-analysis was performed to evaluate the association between GSTM3 and the risk of lung cancer. Methods: Association investigations were identified from PubMed, Embase, and Cochrane Library, and eligible studies were included and synthesized using a meta-analysis method. Results: Eight reports were included into this meta-analysis for the association of GSTM3 A/B gene polymorphism and lung cancer susceptibility, covering 1,854 patients with lung cancer and 1,926 controls. No association between the GSTM3 A/B gene polymorphism and lung cancer was found in this meta-analysis (B allele: OR = 1.25, 95% CI: 0.89-1.76, P = 0.20; BB genotype: OR = 1.53, 95% CI: 0.71-3.32, P = 0.28; AA genotype: OR = 0.85, 95% CI: 0.59-1.23, P = 0.39). Conclusions: The GSTM3 A/B gene polymorphism is not associated with lung cancer susceptibility. However, more studies on the relationship between GSTM3 A/B gene polymorphism and the risk of lung cancer should be performed in the future.

Enhancement of Anticancer Activities of Kimchi by Manipulating Ingredients

  • Kim, Ju-Youn;Rhee, Sook-Hee;Park, Kun-Young
    • Preventive Nutrition and Food Science
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    • 제5권3호
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    • pp.126-130
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    • 2000
  • To enhance the antitumor activity of Chinese cabbage kimchi, four kinds of kimchi, which ere differently prepared in kinds and levels of sub-ingredients, were fermented at 15$^{\circ}C$ for 1 day and then at 5$^{\circ}C$ up to pH 4.3. The solid tumor formation, hepatic glutathione S-transferase activity and glutathione contents in the liver, and natural killer (NK) cell activity of spleen were determined from the sarcoma-180 cell injected Balb/c mice that were treated with methanol extracts of the kimchi samples. Kimchi IV, prepared with organically cultivated Chinese cabbage, red pepper powder, garlic, Chinese pepper powder mustard leaf and heat processed salt (Gueun salt), reduced the tumor formation by 39.3% compared to the sarcoma-180 cell treated group, resulting in the smallest tumor weight. Methanol extracts of the kimchi III and kimchi IV recovered the activities of hepatic glutathione S-transferase(GST) that was decreased by the transplantation of the sarcoma-180 cells to th mice. The injections of methanol extracts of kimchi II and kimchi IV increased glutathione contents in sarcoma-180 cells treated mice. The methanol extract of kimchi IV increased the natural killer (NK) cell activity of spleen lymphocytes a more effectively (p<0.05) than those the other kimchi samples. These results suggest that the anticancer activities of kimchi can be increased by changing the kinds and levels of sub-ingredients.

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황금이 백서의 간 조직 글루타치온 에스-전이 효고 Ycl/2의 발현 효과 (Inductive Effect of Scutellariae radix on Glutathione S-Transferase Yc1/2 from Rat Liver)

  • 김영숙;김동현;최미정;김성민;박래길;권강범;류도곤;김복량
    • 동의생리병리학회지
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    • 제17권6호
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    • pp.1448-1452
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    • 2003
  • The water extract of Scutellariae radix was treated to rat and the primary culture of hepatocytes, and the expressions of several glutathione S-transferase (GST) isozymes and the activity of GST Yc1/2 were investigated by Western blot and by the use of HPLC. The results were obtained as follows: The water extract of Scutellariae radix did not induce the expressions of cytosolic GST Ya and GST Yp in rat livers. But, the extract increased the expression of cytosolic GST Yc1/2 to 2-4 fold higher than control. The expression of GST Yc1/2 in the primary culture of rat hepatocytes was induced by the water extract of Scutellariae radix in a dose-dependent manner, reaching 21-fold over control with 50 ㎍/㎖ treatment. The induction of the expression of GST Yc1/2 in rat livers increased the formation of AFB₁-glutathione conjugate from AFB₁-8,9-epoxide which was made in the metabolism of AFB₁.

토양으로부터 Chlorothalonil 전환 미생물의 분리 및 특성 (Isolation and Characterization of Chlorothalonil-dissipating Bacteria from Soil.)

  • 이수현;신재호;최준호;박종우;김장억;이인구
    • 한국미생물·생명공학회지
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    • 제32권1호
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    • pp.96-100
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    • 2004
  • 토양 시료를 대상으로 chlorothalonil을 함유한 최소배지에서의 집식배양과 배양 추 HPLC에 의한 잔류분석을 통해 chlorothalonil의 제거 능력이 우수한 균주 Ochrobactrum sp. SH35B를 분리하였다. 분리균 SH35B는 1/10 LB 배지에 함유된 10 ppm의 chlorothalonil을 30시간만에 완전히 제거하였으며, 20 ppm의 chlorothalonil의 경우 30시간 동안 88%를 제거하였다. 분리균의 Glu-SH함량과 glutathione S-transferase활성은 각각 1.33및 62.1 nmol/mg이었으며, 대조균인 E. coli나 B. subtilis 보다 높은 것으로 나타났다. 이상의 결과로부터 chlorothalonil의 전환에 있어서 세포내의 Glu-SH 함량과 glutathione S-transferase 활성 이 중요한 인자로 작용하는 것으로 생각된다.

Glutathione S-transferase M1 Null Genotype and Hepatocellular Carcinoma Susceptibility in China and India: Evidence from an Updated Meta-analysis

  • Liu, Hong-Zhou;Peng, Jie;Peng, Chun-Yan;Yan, Ming;Zheng, Fang
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권12호
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    • pp.4851-4856
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    • 2014
  • Background: Glutathione S-transferase M1 (GSTM1) have been reported to be associated with hepatocellular carcinoma. However, the effect of the GSTMl null genotype was divergent in the literature and we therefore performed the present meta-analysis to explore the relationship in detail. Materials and Metbods: Reported studies were searched from 1990 to March 1, 2014 in PubMed and Wanfang Med Online. The total odds oatio (OR) and 95% CI were calculated and analyzed by Review Manager 5.1 and STATE 12. Results: Total OR was calculated from 26 articles with 3,769 cases and 5,517 controls and the association proved significant (OR [95%CI]=1.50 [1.25, 1.80], P<0.05) in the Chinese population. However, there was no significant association between hepatocellular carcinoma risk among subjects carrying the GSTM1 null genotype (OR [95%CI]=1.20 [0.88-1.64], P=0.24) in subgroups of publication in English and in Indian populations (OR [95%CI]=1.80 [0.80-4.20], P=0.15). Conclusions: The GSTM1 deletion polymorphism might not have a significant effect on the susceptibility of hepatocellular carcinoma overall.