• Title/Summary/Keyword: That-Complementation

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A Forward Genetic Approach for Analyzing the Mechanism of Resistance to the Anti-Cancer Drug, 5-Fluorouracil, Using Caenorhabditis elegans

  • Kim, Seongseop;Shim, Jaegal
    • Molecules and Cells
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    • v.25 no.1
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    • pp.119-123
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    • 2008
  • Pyrimidine antagonists including 5-Fluorouracil (5-FU) have been used in chemotherapy for cancer patients for over 40 years. 5-FU, especially, is a mainstay treatment for colorectal cancer. It is a pro-drug that is converted to the active drug via the nucleic acid biosynthetic pathway. The metabolites of 5-FU inhibit normal RNA and DNA function, and induce apoptosis of cancer cells. One of the major obstacles to successful chemotherapy is the resistance of cancer cells to anti-cancer drugs. Therefore, it is important to elucidate resistance mechanisms to improve the efficacy of chemotherapy. We have used C. elegans as a model system to investigate the mechanism of resistance to 5-FU, which induces germ cell death and inhibits larval development in C. elegans. We screened 5-FU resistant mutants no longer arrested as larvae by 5-FU. We obtained 18 mutants out of 72,000 F1 individuals screened, and mapped them into three complementation groups. We propose that C. elegans could be a useful model system for studying mechanisms of resistance to anti-cancer drugs.

A pheromone mutant of Schizosaccharomyces pombe displays nucleolar fragmentation

  • Jun, Jai-Hyun;Kim, Dae-Myung
    • BMB Reports
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    • v.41 no.3
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    • pp.248-253
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    • 2008
  • Stresses and nutritional starvation are two main external signals for the induction of sex pheromones in the fission yeast Schizosaccharomyces pombe. In an attempt to identify the components involved in transduction of starvation signals, we screened 135 temperature-sensitive (ts) mutants and isolated 6 mutants that induced the pheromone even in the presence of a nitrogen source. These mutants exhibited two distict induction phenotypes: pheromone induction at restrictive but not at permissive temperatures; and pheromone induction at both permissive and restrictive temperatures. The times required for the maximum pheromone induction at the restrictive temperature differed slightly in each mutant. In addition to the pheromone induction phenotype, the ts243 and ts304 mutants exhibited cell-division-cycle defects. The ts304 mutant cells showed an abnormal cytoplasmic DAPI staining pattern. The nucleolus of this mutant seemed to be fragmented, a phenomenon which is typically observed in aged yeast cells. The result of our genetic analysis indicated that the pheromone induction mutants belonged to 6 separate complementation groups. We designated these mutants pws1 to pws6.

A Study on the Embodiment -Factors of the Sense of Place in Interior Space -Focused on residential environment (실내공간에서의 장소성 구현 요소에 관한 연구 -주거공간을 중심으로-)

  • 류호창
    • Korean Institute of Interior Design Journal
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    • no.7
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    • pp.56-63
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    • 1996
  • Architectural works build with a modernistic viewpoint led to standardization and low of peculiarity , and which resulted in isolation of people from their environment. In that context, the sense of place can be a new paradigm replacing modernism and reflect the fundamental value and meaning of architecture. Interior space as a place can be analyzed from the viewpoint of existentialism. The purpose of this research is to define the concept of place based on C. N. Schulz's theory of space, and aims at embodiment of the sense of place in interior, especially residential , environment through applying emotional and psychological factors. Spatial characteristics that help to form and enhance the sense of place are center, axis and direction, and boundary and enclosure. Some psychological factors, in other words, people's psychological needs, affect the continuation of the sense of place, especially in residential environment where we reside comparatively for a long period and use the same place repeatedly , security, hierarchy/order, aesthetics, variety, and sociability. The sense of place is embodied and lasted mainly by personalization through the process of visualization , symbolization , and complementation of above mentioned properties and elements.

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Overproduction and Operator DNA-Protein Blotting of R100 Mutant MerR from Shigella flexneri

  • Yoon, Kyung-Pyo
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.250-255
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    • 1994
  • Wild-type and four mutant R100 merR genes were cloned and the proteins overproduced under tac promoter control of pKK223-3. His118Ala, Cys117Ser, Cys126Ser, and wild-type MerR were successfully overproduced although amino-terminal 14 amino acids deletion mutant MerR was not successful. The amount of overproduced wild-type MerR protein as well as other mutant MerR was between 15%-20% of the total protein. The protein was able to be purified up to 95% homogeneity. Specific DNA-protein blotting experiments showed that the 95 bp operator containing DNA fragment could bind to Cys126Ser, His118Ala, and wild- type MerR, but not to Cys117Ser. These results were consistent with the previously reported complementation experiment results that His118Ala, Cys126Ser, and wild-type MerR could repress the mer operon but Cys117Ser could not.

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Expression of a Functional Type-I Chalcone Isomerase Gene Is Localized to the Infected Cells of Root Nodules of Elaeagnus umbellata

  • Kim, Ho Bang;Bae, Ju Hee;Lim, Jung Dae;Yu, Chang Yeon;An, Chung Sun
    • Molecules and Cells
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    • v.23 no.3
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    • pp.405-409
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    • 2007
  • A putative type-I chalcone isomerase (CHI) cDNA clone EuNOD-CHI was previously isolated from the root nodule of Elaeagnus umbellata [Kim et al. (2003)]. To see if it encodes a functional CHI, we ectopically overexpressed it in the Arabidopsis (Arabidopsis thaliana) transparent testa 5 (tt5) mutant, which is defective in naringenin production and has yellow seeds due to proanthocyanidin deficiency. Ectopic overexpression of EuNOD-CHI resulted in recovery of normal seed coat color. Naringenin produced by CHI from naringenin chalcone was detected in the transgenic lines like in the wild-type, whereas it was absent from the tt5 mutant. We conclude that EuNOD-CHI encodes a functional type-I CHI. In situ hybridization revealed that EuNOD-CHI expression is localized to the infected cells of the fixation zone in root nodules.

Glucose Transport through N-Acetylgalactosamine Phosphotransferase System in Escherichia coli C Strain

  • Kim, Hyun Ju;Jeong, Haeyoung;Lee, Sang Jun
    • Journal of Microbiology and Biotechnology
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    • v.32 no.8
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    • pp.1047-1053
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    • 2022
  • When ptsG, a glucose-specific phosphotransferase system (PTS) component, is deleted in Escherichia coli, growth can be severely poor because of the lack of efficient glucose transport. We discovered a new PTS transport system that could transport glucose through the growth-coupled experimental evolution of ptsG-deficient E. coli C strain under anaerobic conditions. Genome sequencing revealed mutations in agaR, which encodes a repressor of N-acetylgalactosamine (Aga) PTS expression in evolved progeny strains. RT-qPCR analysis showed that the expression of Aga PTS gene increased because of the loss-of-function of agaR. We confirmed the efficient Aga PTS-mediated glucose uptake by genetic complementation and anaerobic fermentation. We discussed the discovery of new glucose transporter in terms of different genetic backgrounds of E. coli strains, and the relationship between the pattern of mixed-acids fermentation and glucose transport rate.

An Analysis of Content and Convergence Method of Scientific Technology and Humanities in Elementary School STEAM Programs (과학기술과 인문학 융합 내용 및 융합 방법 실태 분석 -초등학교 융합인재교육(STEAM) 프로그램 개발 과제를 중심으로-)

  • Han, Hyojeong;Kwon, Soonhee
    • Journal of The Korean Association For Science Education
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    • v.37 no.2
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    • pp.313-322
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    • 2017
  • This study aims to analyze the content and methodological aspects of converging scientific technology and humanities in 233 elementary school STEAM programs developed and publicized from 2012 to 2015, to provide the basic materials and implications for editing and supplementing the development of future STEAM programs or policies, or on-site applications of developed programs. The analysis results show that among the Liberal Arts courses of the STEAM program, including Korean, Social Studies, and Ethics, the level of convergence was lowest in Ethics. This seems to be due to the fact that convergence between scientific technology and humanities tended to emphasize a cross-studies convergence over a content convergence done with a specific purpose. In terms of methodological convergence between scientific technology and humanities, the level of convergence of humanities at a normative level was relatively lower than that at a descriptive level. Although a quantitative comparison of the two methods was not significant, it is rather concerning that due to simply converging descriptive humanities, students may not be so aware of the double-sidedness of scientific technology, despite the discovery of some programs that need re-evaluations of their ethical or normative directions. Moreover, the results showed that the cases of converging scientific technology and humanities at a normative level that were analyzed in the STEAM program can be divided into 'complementation of humanities to utilizing scientific technology' and 'complementation of humanities to research in scientific technology and development.' Based on such research results, the study discusses the development of STEAM programs and policies and directions for editing and supplementing programs on-site.

Molecular Cloning, Characterization and Functional Analysis of a 2C-methyl-D-erythritol 2, 4-cyclodiphosphate Synthase Gene from Ginkgo biloba

  • Gao, Shi;Lin, Juan;Liu, Xuefen;Deng, Zhongxiang;Li, Yingjun;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.39 no.5
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    • pp.502-510
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    • 2006
  • 2C-methyl-D-erythritol 2, 4-cyclodiphosphate synthase (MECPS, EC: 4.6.1.12) is the fifth enzyme of the non-mevalonate terpenoid pathway for isopentenyl diphosphate biosynthesis and is involved in the methylerythritol phosphate (MEP) pathway for ginkgolide biosynthesis. The full-length mecps cDNA sequence (designated as Gbmecps) was cloned and characterized for the first time from gymnosperm plant species, Ginkgo biloba, using RACE (rapid amplification of cDNA ends) technique. The full-length cDNA of Gbmecps was 874 bp containing a 720 bp open reading frame (ORF) encoding a peptide of 239 amino acids with a calculated molecular mass of 26.03 kDa and an isoelectric point of 8.83. Comparative and bioinformatic analyses revealed that GbMECPS showed extensive homology with MECPSs from other species and contained conserved residues owned by the MECPS protein family. Phylogenetic analysis indicated that GbMECPS was more ancient than other plant MECPSs. Tissue expression pattern analysis indicated that GbMECPS expressed the highest in roots, followed by in leaves, and the lowest in seeds. The color complementation assay indicated that GbMECPS could accelerate the accumulation of $\beta$-carotene. The cloning, characterization and functional analysis of GbMECPS will be helpful to understand more about the role of MECPS involved in the ginkgolides biosynthesis at the molecular level.

Cloning and Overexpression of the Cdd Gene Encoding Cytidine Deaminase from Salmonella typhimurium

  • Lee, Sang-Mahn
    • Korean Journal of Environmental Biology
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    • v.21 no.1
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    • pp.56-59
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    • 2003
  • The Salmonella typhimurium cdd gene encoding cytidine deaminase (cyti-dine/2'-deoxycytidine aminohydrolase; EC 3.5.4.5.) was isolated through shotgun clon-ing by complementation of the E. coli odd mutation. By subsequent deletion and sub-cloning from the original 3.7 Kb of EcoRI insert (pSAMI), the precise region of the cdd structural gene is located around the BglII site in the middle part of 1.7 Kb of NruI/PvuI segment. The 1.7 Kb containing odd gene wag subcloned to the pUC18 vector and the nucleotide sequence of the cdd gene was determined. When the putative ribosorne-binding site (Shine-Dalgarno sequence) and initiation codon were predicted to be GAGG at the position 459 and ATG at the position 470, respectively, there was an open reading frame of 885 nucleotides, encoding an 294 amino acid protein. The cdd gene expression in E. coli JF611/pSAMI was amplified about 50 fold compared to that of the wild type. The cdd gene expression was maintained in the stationary phase after rea-ching the peak in the late logarithmic phase.

Vegetative Compatibility Groups and Virulence Variation Among Isolates of Pyrenophora graminea

  • Arabi, Mohammad Imad Eddin;Jawhar, Mohammad
    • The Plant Pathology Journal
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    • v.27 no.2
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    • pp.116-119
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    • 2011
  • Pyrenophora graminea, the causal agent of leaf stripe disease, is an economically important pathogen of barley found worldwide. Forty-four isolates of diverse geographical origin within Syria were grouped into vegetative compatibility groups (VCGs) by demonstrating heterokaryosis by complementation tests using nitrate nonutilizing (nit) mutants. All isolates were grouped into three VCGs-1-A, 1-B and 1-C. No self-incompatibility was observed in any of the isolates tested. VCG 1-A was the most common group within growing regions in Syria and proved to be the most virulent of the VCGs identified. These data indicate that the level of virulence in P. graminea is related to VCG.