Updated on 2021/08/31

写真b

 
NOZAKI Shingo
 
*Items subject to periodic update by Rikkyo University (The rest are reprinted from information registered on researchmap.)
Affiliation*
College of Science
Title*
Assistant Professor
Degree
博士(理学) ( 名古屋大学 )
Campus Career*
  • 7 2019 - Present 
    College of Science   Assistant Professor
 

Research Areas

  • Life Science / Applied microbiology  / 合成生物学

Papers

  • Exonuclease III (XthA) enforces in vivo DNA cloning of Escherichia coli to create cohesive ends Peer-reviewed

    Nozaki S, Niki H

    Journal of Bacteriology201 ( 5 )   2 2019

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  • The Ras1-Cdc42 pathway is involved in hyphal development of Schizosaccharomyces japonicus Peer-reviewed

    Nozaki S, Furuya K, Niki H

    FEMS Yeast Research18 ( 4 )   3 2018

  • The Mechanism of Regulation of Pantothenate Biosynthesis by the Pan-DPanZ center dot AcCoA Complex Reveals an Additional Mode of Action for the Antimetabolite N-Pentyl Pantothenamide (N5-Pan) Peer-reviewed

    Zoe L. P. Arnott, Shingo Nozaki, Diana C. F. Monteiro, Holly E. Morgan, Arwen R. Pearson, Hironori Niki, Michael E. Webb

    BIOCHEMISTRY56 ( 37 ) 4931 - 4939   9 2017

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER CHEMICAL SOC  

    The antimetabolite pentyl pantothenamide has broad spectrum antibiotic activity but exhibits enhanced activity against Escherichia coli. The PanDZ complex has been proposed to regulate the pantothenate biosynthetic pathway in E. coli by limiting the supply of beta-alanine in response to coenzyme A concentration. We show that formation of such a complex between activated aspartate decarboxylase (PanD) and PanZ leads to sequestration of the pyruvoyl cofactor as a ketone hydrate and demonstrate that both PanZ overexpression,linked beta-alanine auxottophy and pentyl pantotheriamide toxicity are due to formation of this complex. This both demonstrates that the PanDZ complex regulates pantothenate biosynthesis in a cellular context and validates the complex as a target for antibiotic development.

    DOI: 10.1021/acs.biochem.7b00509

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  • The Structure of the PanD/PanZ Protein Complex Reveals Negative Feedback Regulation of Pantothenate Biosynthesis by Coenzyme A Peer-reviewed

    Diana C. F. Monteiro, Vijay Patel, Christopher P. Bartlett, Shingo Nozaki, Thomas D. Grant, James A. Gowdy, Gary S. Thompson, Arnout P. Kalverda, Edward H. Snell, Hironori Niki, Arwen R. Pearson, Michael E. Webb

    CHEMISTRY & BIOLOGY22 ( 4 ) 492 - 503   4 2015

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:CELL PRESS  

    Coenzyme A (CoA) is an ubiquitous and essential cofactor, synthesized from the precursor pantothenate. Vitamin biosynthetic pathways are normally tightly regulated, including the pathway from pantothenate to CoA. However, no regulation of pantothenate biosynthesis has been identified. We have recently described an additional component in the pantothenate biosynthetic pathway, PanZ, which promotes the activation of the zymogen, PanD, to form aspartate a-decarboxylase (ADC) in a CoA-dependent manner. Here we report the structure of PanZ in complex with PanD, which reveals the structural basis for the CoA dependence of this interaction and activation. In addition, we show that PanZ acts as a CoA-dependent inhibitor of ADC catalysis. This inhibitory effect can effectively regulate the biosynthetic pathway to pantothenate, and thereby also regulate CoA biosynthesis. This represents a previously unobserved mode of metabolic regulation whereby a cofactor-utilizing protein negatively regulates the biosynthesis of the same cofactor.

    DOI: 10.1016/j.chembiol.2015.03.017

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  • Synchronous activation of cell division by light or temperature stimuli in the dimorphic yeast Schizosaccharomyces japonicus Peer-reviewed

    Sho Okamoto, Kanji Furuya, Shingo Nozaki, Keita Aoki, Hironori Niki

    Eukaryotic Cell12 ( 9 ) 1235 - 1243   9 2013

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    Language:English   Publishing type:Research paper (scientific journal)  

    Many fungi respond to light and regulate fungal development and behavior. A blue light-activated complex has been identified in Neurospora crassa as the product of the wc-1 and wc-2 genes. Orthologs of WC-1 and WC-2 have hitherto been found only in filamentous fungi and not in yeast, with the exception of the basidiomycete pathogenic yeast Cryptococcus. Here, we report that the fission yeast Schizosaccharomyces japonicus responds to blue light depending on Wcs1 and Wcs2, orthologs of components of the WC complex. Surprisingly, those of ascomycete S. japonicus are more closely related to those of the basidiomycete. S. japonicus reversibly changes from yeast to hyphae in response to environmental stresses. After incubation at 30°C, a colony of yeast was formed, and then hyphal cells extended from the periphery of the colony. When light cycles were applied, distinct dark- and bright-colored hyphal cell stripes were formed because the growing hyphal cells had synchronously activated cytokinesis. In addition, temperature cycles of 30°C for 12 h and 35°C for 12 h or of 25°C for 12 h and 30°C for 12 h during incubation in the dark induced a response in the hyphal cells similar to that of light. The stripe formation of the temperature cycles was independent of the wcs genes. Both light and temperature, which are daily external cues, have the same effect on growing hyphal cells. A dual sensing mechanism of external cues allows organisms to adapt to daily changes of environmental alteration. © 2013, American Society for Microbiology. All Rights Reserved.

    DOI: 10.1128/EC.00109-13

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  • An activator for pyruvoyl-dependent L-aspartate alpha-decarboxylase is conserved in a small group of the gamma-proteobacteria including Escherichia coli Peer-reviewed

    Shingo Nozaki, Michael E. Webb, Hironori Niki

    MICROBIOLOGYOPEN1 ( 3 ) 298 - 310   9 2012

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-BLACKWELL  

    In bacteria, beta-alanine is formed via the action of L-aspartate alpha-decarboxylase (PanD) which is one of the small class of pyruvoyl-dependent enzymes. The pyruvoyl cofactor in these enzymes is formed via the intramolecular rearrangement of a serine residue in the peptide backbone leading to chain cleavage and formation of the covalently-bound cofactor from the serine residue. This reaction was previously thought to be uncatalysed. Here we show that in Escherichia coli, PanD is activated by the putative acetyltransferase YhhK, subsequently termed PanZ. Activation of PanD both in vivo and in vitro is PanZ-dependent. PanZ binds to PanD, and we demonstrate that a PanZ(N45A) site-directed mutant is unable to enhance cleavage of the proenzyme PanD despite retaining affinity for PanD. This suggests that the putative acetyltransferases domain of PanZ may be responsible for activation to enhance the processing of PanD. Although panD is conserved among most bacteria, the panZ gene is conserved only in E. coli-related enterobacterial species including Shigella, Salmonella, Klebsiella and Yersinia. These bacteria are found predominantly in the gut flora where pantothenate is abundant and regulation of PanD by PanZ allows these organisms to closely regulate production of beta-alanine and hence pantothenate in response to metabolic demand.

    DOI: 10.1002/mbo3.34

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  • Formation of a heterooctameric complex between aspartate alpha-decarboxylase and its cognate activating factor, PanZ, is CoA-dependent Peer-reviewed

    Diana C. F. Monteiro, Michael D. Rugen, Dale Shepherd, Shingo Nozaki, Hironori Niki, Michael E. Webb

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS426 ( 3 ) 350 - 355   9 2012

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    The existence of a fifth essential protein for pantothenate biosynthesis in some enteric bacteria has recently been reported by Stuecker et al. [10] and Nozaki et al. (in press) [9]. This protein, PanZ, catalyses the activation of the PanD zymogen to form ADC and is essential for prototrophic growth. In this paper, we characterise the interaction of PanZ with coenzyme A and a constitutively inactive mutant of PanD using a combination of isothermal titration calorimetry and mass spectrometry. These approaches reveal that the two proteins interact with nanomolar affinity in a CoA-dependent fashion to form a heterooctameric complex. (c) 2012 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2012.08.084

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  • Replication Initiator DnaA of Escherichia coli Changes Its Assembly Form on the Replication Origin during the Cell Cycle Peer-reviewed

    Shingo Nozaki, Hironori Niki, Tohru Ogawa

    JOURNAL OF BACTERIOLOGY191 ( 15 ) 4807 - 4814   8 2009

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC MICROBIOLOGY  

    DnaA is a replication initiator protein that is conserved among bacteria. It plays a central role in the initiation of DNA replication. In order to monitor its behavior in living Escherichia coli cells, a nonessential portion of the protein was replaced by a fluorescent protein. Such a strain grew normally, and flow cytometry data suggested that the chimeric protein has no substantial loss of the initiator activity. The initiator was distributed all over the nucleoid. Furthermore, a majority of the cells exhibited certain distinct foci that emitted bright fluorescence. These foci colocalized with the replication origin (oriC) region and were brightest during the period spanning the initiation event. In cells that had undergone the initiation, the foci were enriched in less intense ones. In addition, a significant portion of the oriC regions at this cell cycle stage had no colocalized DnaA-enhanced yellow fluorescent protein (EYFP) focus point. It was difficult to distinguish the initiator titration locus (datA) from the oriC region. However, involvement of datA in the initiation control was suggested from the observation that, in Delta datA cells, DnaA-EYFP maximally colocalized with the oriC region earlier in the cell cycle than it did in wild-type cells and oriC concentration was increased.

    DOI: 10.1128/JB.00435-09

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  • Initiator titration complex formed at datA with the aid of IHF regulates replication timing in Escherichia coli Peer-reviewed

    Shingo Nozaki, Yoshitaka Yamada, Tohru Ogawa

    GENES TO CELLS14 ( 3 ) 329 - 341   3 2009

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-BLACKWELL PUBLISHING, INC  

    The initiation of replication in Escherichia coli is negatively controlled by a mechanism referred to as 'initiator titration', a process by which the initiator protein, DnaA, is titrated to newly replicated binding sequences on the chromosome to reduce the initiation potential for replication. Initiator titration occurs predominantly at the datA locus that binds exceptionally large amounts of DnaA molecules to prevent aberrant initiations. We found that this was enabled by integration host factor (IHF). Within datA, there is a consensus IHF recognition sequence between the two DnaA recognition sequences (DnaA boxes) essential for its function. Binding of IHF to this site was demonstrated both in vitro and in vivo. Disruption of the core sequence in the consensus of the IHF-binding resulted in increased origin concentration as observed in Delta datA cells. Furthermore, the number of DnaA molecules bound to datA was reduced in cells carrying a disruption in the IHF-binding core sequence. The IHF-binding site and the essential DnaA boxes had to be located at a proper distance and orientation to maintain the accurate initiation timing. Therefore, IHF is a unique element in the control of replication initiation that acts negatively at datA, while known to act as a positive regulator at oriC.

    DOI: 10.1111/j.1365-2443.2008.01269.x

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  • Determination of the minimum domain II size of Escherichia coli DnaA protein essential for cell viability Peer-reviewed

    Shingo Nozaki, Tohru Ogawa

    MICROBIOLOGY-SGM154 ( 11 ) 3379 - 3384   11 2008

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SOC GENERAL MICROBIOLOGY  

    The DnaA protein is the bacterial initiator of replication at a unique chromosomal site, oriC. It is present in all bacterial species and has a conserved structure with four domains. The structures of domains I and III-IV have been solved recently for some bacterial species, and the molecular process leading to the initiation event has been investigated in detail. On the other hand, domain II appears to have no rigid structure and is assumed to be a flexible linker connecting the N-terminal domain I and the C-terminal domains III-IV. It differs significantly in length and amino acid sequence among bacterial species. Whether or not domain II has any function(s) to initiate replication is unknown. The precise borders at both of its ends as well as its essential portions for cell viability are also unknown. In this study, we introduced systematic deletions into the domain II region on the chromosomal dnaA gene of Escherichia coli and examined their effect on cell physiology. Stretches of 30-36 consecutive amino acid residues could be deleted from various portions between the 78th and the 136th residues without affecting cell viability. We propose that domain II of E coli DnaA is from the 79th to the 135th residues and at least 21-27 residues are required as a spacer to keep domains I and III-IV in the correct positions.

    DOI: 10.1099/mic.0.2008/019745-0

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Presentations

  • バクテリオファージを活用した長鎖DNAアセンブリー

    野崎晋五

    第17回大腸菌研究会  20 8 2021 

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  • 大腸菌ゲノムより見出された形質転換に関与する遺伝子群

    野崎晋五, 仁木宏典

    第14回日本ゲノム微生物学会年会  7 3 2020 

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  • 大腸菌細胞内でのDNAクローニング(iVEC)のメカニズムとその応用

    野崎晋五, 仁木宏典

    第1回日本遺伝学会春季分科会  8 3 2019 

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  • 簡便な大腸菌形質転換法を用いたDNA取り込みに関与する遺伝子群のスクリーニング

    野崎晋五, 仁木宏典

    第15回21世紀大腸菌研究会  25 5 2018 

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  • 大腸菌におけるDNA取り込みに関与する遺伝子群の網羅的探索

    野崎晋五, 仁木宏典

    第12回日本ゲノム微生物学会年会  6 3 2018 

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  • NBRPバイオリソースiVEC: 大腸菌細胞内でのDNAクローニングのメカニズムとさらなる高機能化

    野崎晋五, 仁木宏典

    第24回複製・組換え・修復ワークショップ  28 11 2017 

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  • iVEC: 大腸菌細胞内でのDNAクローニングとそのメカニズム

    野崎晋五, 仁木宏典

    第14回21世紀大腸菌研究会  9 6 2017 

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  • iVEC: 大腸菌細胞内でのDNAクローニング方法

    野崎晋五, 仁木宏典

    第39回日本分子生物学会年会  30 11 2016 

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    Language:Japanese  

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  • iVEC: a simple cloning method by using in vivo recombination of E. coli International conference

    Nozaki, S, Niki, H

    The 10th 3R Symposium  14 11 2016 

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    Language:English  

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  • National BioResource Project (NBRP) E. coli and B. subtilis International conference

    Nozaki S, Niki H

    The 8th ANRRC International Meeting  21 9 2016 

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    Language:English  

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  • Ras-Cdc42経路はジャポニカス分裂酵母の菌糸形成と接合の両方に関与する

    野崎晋五, 古谷寛治, 仁木宏典

    第48回酵母遺伝学フォーラム研究報告会  2 9 2015 

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    Language:Japanese  

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  • Crostalk between the mating and hyphal transition at early steps of the Ras-MAPK signal transduction pathway in Schizosaccharomyces japonicus International conference

    Nozaki S, Furuya, K, Niki H

    21 6 2015 

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    Language:English  

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  • Schizosaccharomyces japonicusの菌糸転換におけるCdc2の役割

    野崎晋五, 仁木宏典

    第37回日本分子生物学会年会  26 11 2014 

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    Presentation type:Poster presentation  

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  • Involvement of the inhibition of Cdc2 activity during the yeast cell cycle in hyphal development of Schizosaccharomyces japonicus International conference

    Nozaki, S, Furuya, K, Niki, H

    The 9th 3R symposium  18 11 2014 

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    Presentation type:Poster presentation  

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  • ジャポニカス分裂酵母の菌糸形成に関与する遺伝子の探索

    野崎晋五, 古谷寛治, 吉川博文, 仁木宏典

    第47回酵母遺伝学フォーラム研究報告会  1 9 2014 

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    Presentation type:Poster presentation  

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  • ジャポニカス分裂酵母ゲノムより得られた菌糸形成に関与する遺伝子群

    野崎晋五, 古谷寛治, 吉川博文, 仁木宏典

    第8回日本ゲノム微生物学会年会  7 3 2014 

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    Presentation type:Poster presentation  

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  • Genetic analysis of mutants that are defective in hyphal development of Schizosaccharomyces japonicus International conference

    Nozaki, S, Furuya, K, Niki, H

    7th International Fission Yeast Meeting  24 6 2013 

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    Presentation type:Poster presentation  

    Venue:London  

    国立遺伝学研究所以外との共同発表あり

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  • Regulation of pantothenate and CoA synthesis by a novel factor, PanZ in Escherichia coli

    11 12 2012 

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    Presentation type:Poster presentation  

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  • 新規因子PanZによるアスパラギン酸-α-脱炭酸酵素の活性制御

    野崎晋五, 仁木宏典

    第9回 21世紀大腸菌研究会  21 6 2012 

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    Presentation type:Oral presentation (general)  

    Venue:長浜  

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  • γプロテオバクテリアに現れたパントテン酸合成の新規制御因子PanZ

    野崎晋五, 仁木宏典

    第6回日本ゲノム微生物学会年会  11 3 2012 

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    Presentation type:Poster presentation  

    Venue:池袋  

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  • 新規因子PanZによるピルボイル酵素PanDの活性制御

    野崎晋五, 仁木宏典

    第8回 21世紀大腸菌研究会  18 5 2011 

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    Presentation type:Oral presentation (general)  

    Venue:南木曽  

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  • Regulation of DnaA assembly at oriC in Escherichia coli International conference

    NOZAKI Shingo

    27 10 2010 

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    Presentation type:Poster presentation  

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  • DnaAタンパク質の細胞内局在変化による染色体複製の開始制御

    野崎晋五, 仁木宏典, 小川徹

    第7回21世紀大腸菌研究会  3 6 2010 

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    Presentation type:Oral presentation (general)  

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  • Subcellular localization of Escherichia coli DnaA protein

    Nozaki, S, Niki, H, Ogawa, T

    EMBO Workshop “Frontiers of prokaryotic cell biology”  8 2009 

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  • Subcellular localization of Escherichia coli DnaA, the initiator of DNA replication

    Nozaki, S, Niki, H, Ogawa, T

    Keystone symposia “DNA replication and recombination”  2 2008 

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Research Projects

  • ボトルシップ法による人工細菌の創出

    JST戦略的創造研究推進事業  さきがけ 

    野崎晋五

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    10 2019 - 3 2023

    Authorship:Principal investigator 

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  • 大腸菌の形質転換におけるDNA取り込みメカニズムの解明

    日本学術振興会  科学研究費助成事業 基盤研究(C) 

    野崎 晋五

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    4 2019 - 3 2022

    Grant number:19K05782

    Grant amount:\4420000 ( Direct Cost: \3400000 、 Indirect Cost:\1020000 )

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