• 제목/요약/키워드: GST

검색결과 927건 처리시간 0.032초

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

  • 김동수;진성일
    • 대한전자공학회논문지SP
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    • 제38권4호
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    • pp.411-421
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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.

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

  • 김동수
    • 대한전자공학회논문지SP
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    • 제38권4호
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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.

Generation and characterization of a monoclonal antibody with high species-specificity to Schistosoma japonicum glutathione S-transferase

  • Kim, Jung-Hwan;Park, Jung-Hyun;Ju, Sung-Kyu;Lee, Myung-Kyu;Kim, Kil Lyong
    • IMMUNE NETWORK
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    • 제1권3호
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    • pp.187-195
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    • 2001
  • The expression of recombinant proteins fused to 26 kDa glutathione S-transferase (GST) extracted from Schistosoma japonicum represents an attractive system for purifiying proteins of interest in a single step using GST-affinity chromatography. In addition, the GST-tag is used conveniently for detecting fused proteins since its high solubility as well as its relatively small size rarely interferes with the biological activity of the fused protein. In this regard, the GST system is frequently applied for tracing fusion proteins in both prokaryotic and eukaryotic cells to elucidate the physiological interactions and functional compartments of proteins. To provide a further tool in analyzing GST-fusion proteins, a new monoclonal antibody, with a high specificity to the S. japonicum GST was produced. Methods: BALB/c mice were immunized both with recombinant S. japonicum GST proteins, and by the fusion of splenocytes from these mice with myeloma cells. From this, a new anti -GST monoclonal antibody, termed SARAH, was generated. The specificity and reactivity of this antibody was confirmed by ELISA and by Western blot analysis. Results: SARAH showed a high reactivity to recombinant GST and GST fusion protein but not with native mammalian GST proteins as derived from other species including humans, cows, rabbits and rats. The applicability of SARAH was further demonstrated by confocal laser scanning microscopy, where GST proteins that were expressed transiently in mouse fibroblast cells, were specifically detected without interference of endogenous GST. Conclusion: SARAH is new monoclonal antibody with a high specificity to recombinant GST proteins but not to endogenous GST in mammalian cells.

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

  • 임태형;박혜선
    • 사상체질의학회지
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    • 제27권2호
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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.

북방전복 (Haliotis discus hannai)에서 분리한 Glutathione S-transferase 유전자의 분자생물학적 고찰 및 발현분석 (Molecular Characterization and Expression Analysis of a Glutathione S-Transferase cDNA from Abalone (Haliotis discus hannai))

  • 문지영;박은희;공희정;김동균;김영옥;김우진;안철민;남보혜
    • 한국패류학회지
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    • 제30권4호
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    • pp.399-408
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    • 2014
  • 본 연구에서는 북방전복 (Haliotis discus hannai)의 대용량 염기서열 분석을 통해 GST유전자의 전장 cDNA를 동정하였다. 북방전복 GST 유전자의 총 길이는 1669 bp로 672 bp의 ORF는 총 223개의 아미노산을 코딩하고 있으며 등전점은 5.69, 분자량은 25.8 kDa으로 예측되었다. 북방전복 GST아미노산 서열은 둥근전복과 지중해 담치와 같은 패류의 GSTA와 가장 유사성이 높았으며 계통수 분석을 통해 GSTA와 하나의 그룹을 이루었다. 북방전복 GST에는 GSTA의 특징을 갖는 두 site (N-말단의 G-site, C-말단의 H-site)가 보존되어 있었고 효소활성과 구조 유지에 중요한 잔기가 종간에 매우 보존되어 있었다. 북방전복 GST 유전자의 mRNA는 관찰된 모든 조직에서 발현하고 있었으며, 특히 외투막, 아가미, 간췌장, 소화관에서 높은 발현이 확인되었다. 북방전복의 GST는 비브리오균을 인위감염 시킨 전복의 간췌장에서 감염 후 1시간 뒤 발현이 급격히 증가했다가 3시간까지 증가한 뒤 감소하였고, 혈구세포에서는 감염 3시간 경과 후 발현 정도가 최고로 증가했다가 감소하였다. 따라서 북방전복 GST는 alpha class GST의 특징을 가지며 병원체 감염에 따른 면역반응 조절에 관여할 것이라 생각되며 병원균 감염에 따른 바이오마커로 활용가능 할 것이라 예상된다.

Characterization of binding specificity using GST-conjugated mutant huntingtin epitopes in surface plasmon resonance (SPR)

  • Cho, Hang-Hee;Kim, Tae Hoon;Kim, Hong-Duck;Cho, Jae-Hyeon
    • 한국동물위생학회지
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    • 제44권4호
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    • pp.185-194
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    • 2021
  • Polyglutamine extension in the coding sequence of mutant huntingtin causes neuronal degeneration associated with the formation of insoluble polyglutamine aggregates in Huntington's disease (HD). Mutant huntingtin can form aggregates within the nucleus and processes of neurons possibly due to misfolding of the proteins. To better understand the mechanism by which an elongated polyglutamine causes aggregates, we have developed an in vitro binding assay system of polyglutamine tract from truncated huntingtin. We made GST-HD exon1 fusion proteins which have expanded polyglutamine epitopes (e.g., 17, 23, 32, 46, 60, 78, 81, and 94 CAG repeats). In the present emergence of new study adjusted nanotechnology on protein chip such as surface plasmon resonance strategy which used to determine the substance which protein binds in drug discovery platform is worth to understand better neurodegenerative diseases (i.e., Alzheimer disease, Parkinson disease and Huntington disease) and its pathogenesis along with development of therapeutic measures. Hence, we used strengths of surface plasmon resonance (SPR) technology which is enabled to examine binding specificity and explore targeted molecular epitope using its electron charged wave pattern in HD pathogenesis utilize conjugated mutant epitope of HD protein and its interaction whether wild type GST-HD interacts with mutant GST-HD with maximum binding affinity at pH 6.85. We found that the maximum binding affinity of GST-HD17 with GST-HD81 was higher than the binding affinities of GST-HD17 with other mutant GST-HD constructs. Furthermore, our finding illustrated that the mutant form of GST-HD60 showed a stronger binding to GST-HD23 or GST-HD17 than GST-HD60 or GST-HD81. These results indicate that the binding affinity of mutant huntingtin does not correlate with the length of polyglutamine. It suggests that the aggregation of an expanded polyglutamine might have easily occurred in the presence of wild type form of huntingtin.

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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    • 제44권4호
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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.

Production and characterization of a monoclonal anti-glutathione-S-transferase(GST) antibody

  • You, Je-Kyung;Shin, Chan-Young;Park, Kyu-Hwan;Ko, Kwang-Ho
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1997년도 춘계학술대회
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    • pp.93-93
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    • 1997
  • Analysis of protein is often frustrated by the inability to isolate large amounts of purified protein from a native source. To overcome this problem, fusion protein expression systems such as pGEX system have been widely used. Using pGEX system, the desired protein could be easily obtained in a large amount in E. coli, and then the fusion protein could be used for the study of the function of the given protein. To analyze and purify the GST fusion protein, anti-GST antibody could be used as one of the system of choice. However, the production and characterization of monoclonal anti-GST antibody has not been studied extensively yet. To produce monoclonal anti-GST antibody, GST was purified from E. coli transformed with pGEX-cs, one of the pGEX system and was used as an antigen. The monoclonal antibody was produced by fusion of the immunized spleen cells with SP2-0 myeloma cells. The antibody was characterized by ELISA, western blotting, etc. The monoclonal antibody produced in this study (mAb-GSTA) showed strong and specific immunoreactivity against not only GST but also GST-fusion proteins. Also, mAb-GSTA was successfully used for the immunoaffinity purification of the GST ${\beta}$-Rc.-third intracellular-loop fusion protein. The results of the present study suggest that mAb-GSTA may be used for the identification and purification of GST fusion 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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    • 제24권9호
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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.