• Title/Summary/Keyword: GST2

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Funcyional Studies on Gene 2.5 Protein of Bacteriophage T7 : Protein Interactions of Replicative Proteins (박테리오파아지 T7 의 기능에 관한 연구;복제단백질간의 단백질 상호작용)

  • 김학준;김영태
    • Journal of Life Science
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    • v.6 no.3
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    • pp.185-192
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    • 1996
  • Bacteriophage T7 gene 2.5 protein, a single-stranded DNA binding protein, is required for T7 DNA replication, recombination, and repair. T7 gene 2.5 protein has two distinctive domains, DNA binding and C-terminal domain, directly involved in protein-protein interaction. Gene 2.5 protein participates in the DNA replication of Bacteriophage T7, which makes this protein essential for the T7 growth and DNA replication. What gene 2.5 protein makes important at T7 growth and DNA replication is its binding affinity to single-stranded DNA and the protein-protein important at T7 DNA replication proteins which are essential for the T7 DNA synthesis. We have constructed pGST2.5(WT) encoding the wild-type gene 2.5 protein and pGST2.5$\Delta $21C lacking C-terminal 21 amino acid residues. The purified GST-fusion proteins, GST2.5(WT) and GST2.5(WT)$\Delta$21C, were used for whether the carboxyl-terminal domain participates in the protein-protein interactions or not. GST2.5(WT) and GST2.5$\Delta$21C showed the difference in the protein-protein interaction. GST2.5(WT) interacted with T7 DNA polymerase and gene 4 protein, but GST2.5$\Delta$21C did not interact with either protein. Secondly, GST2.5(WT) interacts with gene 4 proteins (helicase/primase) but not GST2.5$\Delta$21C. these results proved the involvement of the carboxyl-terminal domain of gene 2.5 protein in the protein-protein interaction. We clearly conclude that carboxy-terminal domain of gene 2.5 protein is firmly involved in protein-protein interactions in T7 replication proteins.

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GST2 is Required for Nitrogen Starvation-Induced Filamentous Growth in Candida albicans

  • Lee, So-Hyoung;Chung, Soon-Chun;Shin, Jongheon;Oh, Ki-Bong
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1207-1215
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    • 2014
  • Candida albicans, the major human fungal pathogen, undergoes morphological transition from the budding yeast form to filamentous growth in response to nitrogen starvation. In this study, we identified a new function of GST2, whose expression was required for filamentous growth of C. albicans under nitrogen-limiting conditions. The Gst2p showed Gst activity and required response to oxidative stress. The ${\Delta}gst2$ mutant displayed predominantly yeast phase growth in low ammonium media. Such morphological defect of ${\Delta}gst2$ mutants was not rescued by overexpression of Mep2p, Cph1p, or Efg1p, but was rescued by either overexpression of a hyperactive $RAS1^{G13V}$ allele or through exogenous addition of cyclic AMP. In addition, the ${\Delta}gst2$ mutants had lower levels of RAS1 transcripts than wild-type cells under conditions of nitrogen starvation. These results were consistent with the Ras1-cAMP pathway as a possible downstream target of Gst2p. These findings suggest that Gst2p is a significant component of nitrogen starvation-induced filamentation in C. albicans.

Improved recovery of active GST-fusion proteins from insoluble aggregates: solubilization and purification conditions using PKM2 and HtrA2 as model proteins

  • Park, Dae-Wook;Kim, Sang-Soo;Nam, Min-Kyung;Kim, Goo-Young;Kim, Jung-Ho;Rhim, Hyang-Shuk
    • BMB Reports
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    • v.44 no.4
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    • pp.279-284
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    • 2011
  • The glutathione S-transferase (GST) system is useful for increasing protein solubility and purifying soluble GST fusion proteins. However, purifying half of the GST fusion proteins is still difficult, because they are virtually insoluble under non-denaturing conditions. To optimize a simple and rapid purification condition for GST-pyruvate kinase muscle 2 (GST-PKM2) protein, we used 1% sarkosyl for lysis and a 1 : 200 ratio of sarkosyl to Triton X-100 (S-T) for purification. We purified the GST-PKM2 protein with a high yield, approximately 5 mg/L culture, which was 33 times higher than that prepared using a conventional method. Notably, the GST-high-temperature requirement A2 (GST-HtrA2) protein, used as a model protein for functional activity, fully maintained its proteolytic activity, even when purified under our S-T condition. This method may be useful to apply to other biologically important proteins that become highly insoluble in the prokaryotic expression system.

Overexpression of the $E1_{192-283}$ and $E2_{384-649}$ Proteins of Hepatitis C Virus in GST Fusion Forms in E. coli and Their Immunogenicity (C 형 간염 바이러스의 외피당단백질 E1 및 E2의 융합단백질 $GST-E1_{192-283}$$-E2_{384-649}$의 대장균에서의 과량발현 및 면역원성 연구)

  • Seong, Young-Rim;Choi, See-Young;Im, Dong-Soo
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.105-113
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    • 1997
  • The truncated $E1_{192-283}$ and $E2_{384-649}$ genes of hepatitis C virus (HCV) linked to the gene for glutathione S-transferase (GST) were constructed and their expressions were analyzed. The $GST-E1_{192-283}$ fusion gene overexpressed the fusion protein in E. coli as a soluble form, while the $GST-E1_{192-383}$ plasmid did not express expected fusion protein. The purified $GST-E1_{192-283}$ fusion protein was efficiently cleaved by thrombin. More than 90% pure, HCV $E1_{192-283}$ protein was obtained by GST-agarose chromatography. The truncated $GST-E2_{384-649}$ fusion gene expressed the fusion protein mainly as an insoluble form, whereas the $GST-E2_{384-740}$ did not express the fusion protein. The truncated $GST-E1_{182-283}$ and $GST-E2_{384-649}$ fusion proteins reacted specifically with an HCV patient serum. In addition, mice immunized with either the purified $E1_{192-283}$ or $GST-E2_{384-649}$ proteins generated specific antibodies to each antigen. The results suggested that hydrophobic carboxyl portions of the E1 and E2 proteins might affect expression levels as well as the solubility of each fusion protein in bacteria. Also, the truncated E1 protein with Tyr-192 to Ser-283 contained antigenic epitope(s) which could be specifically recognized by an HCV patient serum.

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The Effects of GalGunSeunggi-tang (GST) on Mice Induced Allergic Contact Dermatitis (ACD) by DNCB (DNCB로 유발된 생쥐의 알레르기접촉피부염에 갈근승기탕(葛根承氣湯)이 미치는 영향)

  • Lim, Tae-Hyung;Park, Hye-Sun
    • Journal of Sasang Constitutional Medicine
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    • v.27 no.2
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    • pp.240-253
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    • 2015
  • Objectives The aim of this study was to investigate the effects of GalGunSeungGi-Tang (GST) on allergic contact dermatitis (ACD) induced by 2,4-dintrochlorobenzone (DNCB) Methods In this study, The changes of body weight, ear weight, ear thickness, spleen weight, dorsum skin thickness, symptom score by eyesight, histological finding, proliferation rates of splenocyte in vitro and in vivo are investigated to check effects of GST. The mice are divided into four group; Normal (naive mice), Control (DW administered), GST-L (GST 500mg/Kg/day administered), GST-H (GST 1,000mg/Kg/day administered). Results GST inhibited weight of ear significantly (P < 0.05) and also thickness (P < 0.01). In addition, There are significant decrease in thickness of dorsum skin and proliferation rates of splenocyte in vivo in GST administered group. Finally, GST reduced symptom score and hyperkeratosis, hyperpigmentation, increase of granulocyte and parakeratosis in histological finding. Conclusions These results suggest that GST can decrease symptoms of ACD.

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.

Modified Scan Line Based Generalized Symmetry transform with selectively Directional Attention (선택적 방향 주의를 가지는 수정된 스캔 라인 일반화 대칭 변환)

  • Kim, Dong Su
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.38 no.4
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    • pp.87-87
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    • 2001
  • 일반화 대칭 변환 (generalized symmetry transform, GST)은 주어진 영상에서 사전 분할이 없이 국부성과 반사 대칭성을 결합하여 대칭을 측정하고 관심 영역을 추출한다. GST의 거리 가중치 함수에서 국부적인 대칭성이 반영되며 이 함수의 표준 편차 u에 의해 GST의 수행 범위가 조절된다. 넓은 관심영역을 추출하기 위해 반지름 r이 큰 검색영역 내에서의 대칭성이 추출될 필요가 있다. 이에 따라서 GST의 수행시간은 r에 따라 2차적으로 증가하게 된 본 논문에서는 이를 개선하기 위해 선택적 방향 주의를 가지는 수정된 스캔라인 GST를 제안한다. 제안된 GST는 기존의 GST와 유사한 대칭 특성을 추출하지만 선택적 방향의 기울기만을 고려한 스캔라인 위의 에지 화소쌍에서 GST를 수행함으로써 r에 따라서 이의 수행시간이 선형적으로 증가된다 특히 r이 큰 경우에 선택적 방향에 대해서만 적용하면 기존의 GST의 계산량이 비대해지는 단점을 보완해 줄 수 있다. 제안된 GST가 기존의 GST보다 시간적으로 효과적이며 유용하다는 것이 여러 종류의 영상에 대한 실험으로 확인되었다.

실시간 비저항 측정을 통한 N-doped $Ge_2Sb_2Te_5$ 박막의 결정화에 대한 연구

  • Lee, Do-Gyu;Do, Gi-Hun;Son, Hyeon-Cheol;Go, Dae-Hong
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.02a
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    • pp.136-136
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    • 2010
  • $Ge_2Sb_2Te_5$ (GST)는 광학 스토리지 및 PRAM(Phase-change Random Access Memory)에 적용 가능한 대표적인 상변화 물질이며 상변화 거동에 대한 다양한 연구가 진행되고 있다. 차세대 비휘발성 메모리로 각광을 받고 있는 PRAM의 경우 저전력 그러나 향후 고집적, 고성능 PRAM 소자구현을 위해서는 Reset 전류 감소를 통한 소비 전력 감소, 인접 셀간의 'cross talking'을 방지할 수 있는 열적 안정성 개선 등의 문제점들을 해결해야 한다. GST 물질의 전기적, 열적 특성을 조절하여 이러한 문제를 해결하기 위하여 GST 물질에 이종의 원소를 첨가하는 연구가 활발히 진행되고 있으며, 특히 질소 첨가에 의해 결정 성장 억제를 통한 결정화 온도 증가, 결정질의 저항 증가 등의 보고가 있었다. 본 연구에서는 질소를 첨가한 N-doped $Ge_2Sb_2Te_5$ (NGST) 박막의 상변화 거동을 규명하고 GST 박막과 비교하여 첨가된 질소의 영향을 분석하고자 한다. D.C Magnetron sputtering 방법으로 증착된 GST와 NGST 박막을 등온으로 유지하여 각 온도별로 열처리 시간 증가에 따른 비저항을 실시간으로 측정하여 GST와 NGST 박막의 상분율을 계산하고 Kissinger 모델을 이용하여 effective activation energy ($E_a$)를 구하였다. GST와 NGST 박막의 $E_a$는 각각 $2.08\;{\pm}\;0.11\;eV$$2.66\;{\pm}\;0.12\;eV$로 계산되었다. 따라서 첨가된 질소에 의해 NGST 박막의 결정화를 위하여 GST 박막의 경우보다 더 큰 활성화 에너지가 필요하다.

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The Expression of Hsp70 and GST Genes in Mytilus coruscus during Air Exposure and Starvation (공기노출 및 절식시기 동안 참담치, Mytilus coruscus 에서 Hsp70 및 GST 유전자 발현에 대한 연구)

  • Kim, Chul Won;Kang, Han Seung
    • The Korean Journal of Malacology
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    • v.32 no.2
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    • pp.73-81
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    • 2016
  • Heat shock proteins (HSPs), one of the most highly conserved groups of proteins characterized to date, play crucial roles in protecting cells against environmental stresses, such as heat shock, salinity and oxidative stress. The glutathione S-transferases (GST) have important role in detoxification of oxidative stress, environmental chemicals and environmental stress. GST mRNA expression have been used as biomarkers on environmental stress. The purpose of this study was to investigate the death rate and the gene expression of Hsp70 and GST during air exposure and starvation. Results showed that, the expression of Hsp70 mRNA was significantly changed in the experiment groups, such as air exposure and starvation. GST mRNA expression was significantly increased in the experimental group of starvation. These results suggest that Hsp70 and GST were played roles in biomarker gene on the air exposure and starvation.

Modified Scan Line Based Generalized Symmetry Transform with Selectively Directional Attention (선택적 방향주의를 가지는 수정된 스캔라인 일반화 대칭 변환)

  • Kim, Dong-Su;Jin, Seong-Il
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.38 no.4
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    • pp.411-421
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    • 2001
  • The generalized symmetry transform evaluates symmetry without segmentation and extracts regions of interest in an image by combining locality and reflectional symmetry The demand that the symmetry transform be local is reflected by the distance weight function. When calculating large regions-of-interest, we should select a large standard deviation of distance weight function. But such a large standard deviation makes the execution time increase in the second power of r, which is a radius of search area. In this paper we propose modified scan line based GST with selectively directional attention to improve time complexity The symmetry map of our proposed GST is found to be very similar to that of the existing GST. However the computation time of the proposed GST increases linearly with respect to r because our proposed GST evaluates symmetry between a pair of edge pixels along the scan lines. The GST computation decreases considerably when the proposed GST is peformed with selectively directional attention in case of large r. Several experiments in this paper demonstrate the time efficiency and the usefulness of our proposed GST.

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