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Creation of stress tolerant strain based on single nucleotide mutational analysis of in whole genome

Research Project

Project/Area Number 21360401
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Biofunction/Bioprocess
Research InstitutionOsaka University

Principal Investigator

SHIMIZU Hiroshi  大阪大学, 大学院・情報科学研究科, 教授 (00226250)

Co-Investigator(Kenkyū-buntansha) FURUSAWA Chikara  大阪大学, 大学院・情報科学研究科, 准教授 (00372631)
HIRASAWA Takashi  大阪大学, 大学院・情報科学研究科, 助教 (20407125)
ONO Naoaki  大阪大学, 大学院・情報科学研究科, 特任准 (60395118)
Project Period (FY) 2009 – 2011
Project Status Completed (Fiscal Year 2011)
Budget Amount *help
¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000)
Fiscal Year 2011: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
Fiscal Year 2010: ¥7,150,000 (Direct Cost: ¥5,500,000、Indirect Cost: ¥1,650,000)
Fiscal Year 2009: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
Keywordsゲノム / DNAマイクロアレイ / 変異解析 / ストレス耐性 / 進化工学 / バイオプロダクション / 発現プロファイル / 比増殖速度
Research Abstract

We performed experimental evolution under(5%) ethanol stress condition in Escherichia coli and six ethanol stress tolerant evolved strains were independently obtained. Transcriptomic analysis of evolved strains was performed. As a result of principal component analysis(PCA), gene expression in biosynthesis pathways of some amino acids were extracted and these are commonly up-regulated in the tolerant strains, suggesting that activating these pathways were involved in the achievement of ethanol tolerance. In support of this hypothesis, the supplementation of these amino acids to the culture medium increased the specific growth rate under ethanol stress. Mutational analysis was also performed by high-density oligonucleotide microarray and it was suggested that mutation does not significantly contribute to ethanol stress tolerance because there few common mutations among evolved strains.

Report

(4 results)
  • 2011 Annual Research Report   Final Research Report ( PDF )
  • 2010 Annual Research Report
  • 2009 Annual Research Report
  • Research Products

    (58 results)

All 2012 2011 2010 2009 Other

All Journal Article (9 results) (of which Peer Reviewed: 8 results) Presentation (43 results) Book (2 results) Remarks (4 results)

  • [Journal Article] Transcriptome analysis of parallel-evolved Escherichia coli strains under ethanol stress2010

    • Author(s)
      Takaaki Horinouchi, Kuniyasu Tamaoka, Chikara Furusawa, Naoaki Ono, Shingo Suzuki, Takashi Hirasawa, Tetsuya Yomo, Hiroshi Shimizu.
    • Journal Title

      BMC Genomics

      Volume: 11 Issue: 1 Pages: 579-579

    • DOI

      10.1186/1471-2164-11-579

    • Related Report
      2011 Final Research Report
  • [Journal Article] Saccharomyces cerevisiae and DNA microarray analyses : what did we learn from it for a better understanding and exploitation of yeast biotechnology2010

    • Author(s)
      Takashi Hirasawa
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: Vol.87 Pages: 391-400

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Transcriptome analysis of parallel-evolved Escherichia coli strains under ethanol stress2010

    • Author(s)
      Takaaki Horinouchi, Hiroshi Shimizu
    • Journal Title

      BMC Genomics

      Volume: Vol.11 Pages: 579-579

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Relationship between noise characteristics in protein expressions and regulatory structures of amino acid biosynthesis pathways2010

    • Author(s)
      Tadashi Yamada
    • Journal Title

      IET Systems Biology 4

      Pages: 82-89

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Differential importance of trehalose accumulation in Saccharomyces cerevisiae in response to various environmental stresses2010

    • Author(s)
      Siraje Arif Mahmud
    • Journal Title

      Journal of Bioscience and Bioengineering 109

      Pages: 262-266

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Requirement of de novo synthesis of the OdhI protein in penicillin-induced glutamate productioh by Corynebacterium glutamicum2010

    • Author(s)
      Jongpill Kim
    • Journal Title

      Applied Microbiology and Biotechnology 印刷中(印刷中)

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Developement and experimental verification of a genome-scale metabolicmodel for Corynebacterium glutamicum2009

    • Author(s)
      Yohei Shinfuku
    • Journal Title

      Microbial Cell Factory 8

      Pages: 43-47

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Investigating the effectiveness of DNA microarray analysis for identifying the genes involved in L-lactate production by Saccharomyces cerevisiae2009

    • Author(s)
      Takashi Hirasawa
    • Journal Title

      Applied Microbiology and Biotechnology 84

      Pages: 1149-1159

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Analysis of stochasticity in promoter activation by using a dual-fluorescence reporter system2009

    • Author(s)
      Jianhong Ou
    • Journal Title

      BioSystems 97

      Pages: 160-164

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Presentation] Integration of in silico and experimental approaches for creation of microbial cell factories2012

    • Author(s)
      Hiroshi Shimizu
    • Organizer
      14th Asia Pacific Confederation of Chemical Engineering(APCChE 2012)
    • Place of Presentation
      Suntec Singapore International Convention & Exhibition Center, Singapore
    • Year and Date
      2012-02-24
    • Related Report
      2011 Final Research Report
  • [Presentation] Integration of in silico and experimental approaches for creation of microbial cell factories2012

    • Author(s)
      Hiroshi Shimizu
    • Organizer
      14th Asia Pacific Confederation of Chemical Engineering (APCChE 2012)
    • Place of Presentation
      Suntec Singapore International Convention & Exhibition Center (Singapore)(招待講演)
    • Year and Date
      2012-02-24
    • Related Report
      2011 Annual Research Report
  • [Presentation] 大腸菌エタノール耐性進化株の次世代シーケンサを用いた全ゲノム解析2011

    • Author(s)
      小野直亮, 堀之内貴明, 玉岡邦康, 鈴木真吾, 平沢敬, 古澤力, 四方哲也, 清水浩
    • Organizer
      第64回日本生物工学会大会
    • Place of Presentation
      東京農工大学
    • Year and Date
      2011-09-28
    • Related Report
      2011 Final Research Report
  • [Presentation] 大腸菌エタノール耐性進化株の次世代シーケンサを用いた全ゲノム解析2011

    • Author(s)
      小野直亮
    • Organizer
      第64回日本生物工学会大会
    • Place of Presentation
      東京農工大学(東京都)
    • Year and Date
      2011-09-28
    • Related Report
      2011 Annual Research Report
  • [Presentation] Whole genome resequencing of experimentally evolved Escherichia coli using a next generation sequencer2011

    • Author(s)
      小野直亮, 堀之内貴明, 玉岡邦康, 鈴木真吾, 平沢敬, 古澤力, 四方哲也, 清水浩
    • Organizer
      日本生物物理学会第49回年会
    • Place of Presentation
      兵庫県立大学
    • Year and Date
      2011-09-16
    • Related Report
      2011 Final Research Report
  • [Presentation] Whole genome resequencing of experimentally evolved Escherichia coli using a next generation sequencer2011

    • Author(s)
      小野直亮
    • Organizer
      日本生物物理学会第49回年会
    • Place of Presentation
      兵庫県立大学(兵庫県)
    • Year and Date
      2011-09-16
    • Related Report
      2011 Annual Research Report
  • [Presentation] エタノールストレス環境下の進化実験により得られた大腸菌のゲノムワイドな変異解析と遺伝子発現解析2011

    • Author(s)
      清水浩
    • Organizer
      化学工学会第43回秋季大会
    • Place of Presentation
      名古屋工業大学(愛知県)
    • Year and Date
      2011-09-15
    • Related Report
      2011 Annual Research Report
  • [Presentation] エタノールストレス環境下の進化実験により得られた大腸菌のゲノムワイドな変異解析と遺伝子発現解2011

    • Author(s)
      堀之内貴明, 玉岡邦康, 古澤力, 小野直亮, 鈴木真吾, 平沢敬, 四方哲也, 清水浩
    • Organizer
      化学工学会第43回秋季大会
    • Place of Presentation
      名古屋工業大学
    • Year and Date
      2011-09-05
    • Related Report
      2011 Final Research Report
  • [Presentation] illuminaを用いた大腸菌全ゲノムリシーケンシング2011

    • Author(s)
      小野直亮, Roche, SOLiD
    • Organizer
      NGS(次世代シークエンサ)現場の会第1回研究会
    • Place of Presentation
      熱海ニューフジヤホテル,静岡県
    • Year and Date
      2011-05-29
    • Related Report
      2011 Final Research Report
  • [Presentation] Roche, SOLiD, illuminaを用いた大腸菌全ゲノムリシーケンシング2011

    • Author(s)
      小野直亮
    • Organizer
      NGS(次世代シークエンサ)現場の会第1回研究会
    • Place of Presentation
      熱海ニューフジヤホテル(静岡県)
    • Year and Date
      2011-05-29
    • Related Report
      2011 Annual Research Report
  • [Presentation] Genome-Wide Mutational and Expression Analyses of Evolved Escherichia Coli Strains under Ethanol Stress2011

    • Author(s)
      Naoaki Ono, Chikara Furusawa, Takaaki Horinouchi, Kuniyasu Tamaoka, Shingo Suzuki, Takashi Hirasawa, Tetsuya Yomo Hiroshi Shimizu
    • Organizer
      Asian Congress on Biotechnology(ACB-2011)
    • Place of Presentation
      Shanghai, China
    • Year and Date
      2011-05-13
    • Related Report
      2011 Final Research Report
  • [Presentation] Genome-Wide Mutational and Expression Analyses of Evolved Escherichia Coli Strains under Ethanol Stress2011

    • Author(s)
      Naoaki Ono
    • Organizer
      Asian Congress on Biotechnology (ACB-2011)
    • Place of Presentation
      Shanghai Everbright Convention & Exhibition Centre (SECEC), Shanghai (China)
    • Year and Date
      2011-05-13
    • Related Report
      2011 Annual Research Report
  • [Presentation] 人工進化工学と網羅的遺伝子発現情報解析によるストレス耐性細胞の創製2011

    • Author(s)
      堀之内貴明, 玉岡邦康, 古澤力, 小野直亮, 鈴木真吾, 平沢敬, 四方哲也, 清水浩
    • Organizer
      化学工学会第76年会
    • Place of Presentation
      東京農工大学
    • Year and Date
      2011-03-23
    • Related Report
      2011 Final Research Report
  • [Presentation] 人工進化工学と網羅的遺伝子発現情報解析によるストレス耐性細胞の創製2011

    • Author(s)
      清水浩
    • Organizer
      化学工学会 第76年会
    • Place of Presentation
      東京農工大学小金井キャンパス、東京都
    • Year and Date
      2011-03-23
    • Related Report
      2010 Annual Research Report
  • [Presentation] エタノール環境下での長期植え継ぎ培養によって得られた耐性大腸菌株の遺伝子発現・ゲノム変異解析2010

    • Author(s)
      堀之内貴明, 玉岡邦康, 古澤力, 小野直亮, 鈴木真吾, 平沢敬, 四方哲也, 清水浩
    • Organizer
      第33回日本分子生物学会年会・第83回日本生化学会大会合同大会(BMB2010)
    • Place of Presentation
      神戸ポートピアアイランド,兵庫県
    • Year and Date
      2010-12-10
    • Related Report
      2011 Final Research Report
  • [Presentation] エタノール環境下での長期植え継ぎ培養によって得られた耐性大腸菌株の遺伝子発現・ゲノム変異解析2010

    • Author(s)
      古澤力
    • Organizer
      第33回日本分子生物学会年会・第83回日本生化学会大会合同大会(BMB2010)
    • Place of Presentation
      神戸ポートピアアイランド、兵庫県
    • Year and Date
      2010-12-10
    • Related Report
      2010 Annual Research Report
  • [Presentation] 人工進化実験における発現制御ダイナミクスの進化的シミュレーション2010

    • Author(s)
      堀之内貴明, 古澤力, 清水浩, 四方哲也, 小野直亮
    • Organizer
      定量生物学の会第三回年会
    • Place of Presentation
      東京大学生産技術研究所
    • Year and Date
      2010-11-27
    • Related Report
      2011 Final Research Report
  • [Presentation] 大腸菌エタノールストレス人工進化株の網羅的解析によるエタノール耐性付与2010

    • Author(s)
      堀之内貴明, 玉岡邦康, 古澤力, 平沢敬, 小野直亮, 鈴木真吾, 四方哲也, 清水浩
    • Organizer
      定量生物学の会第三回年会
    • Place of Presentation
      東京大学生産技術研究所
    • Year and Date
      2010-11-27
    • Related Report
      2011 Final Research Report
  • [Presentation] 大腸菌エタノールストレス人工進化株の網羅的解析によるエタノール耐性付与2010

    • Author(s)
      堀之内貴明、清水浩
    • Organizer
      定量生物学の会 第三回年会
    • Place of Presentation
      東京大学生産技術研究所、東京都
    • Year and Date
      2010-11-27
    • Related Report
      2010 Annual Research Report
  • [Presentation] 人工進化実験における発現制御ダイナミクスの進化的シミュレーション2010

    • Author(s)
      小野直亮
    • Organizer
      定量生物学の会 第三回年会
    • Place of Presentation
      東京大学生産技術研究所、東京都
    • Year and Date
      2010-11-27
    • Related Report
      2010 Annual Research Report
  • [Presentation] 次世代シーケンサを用いた大腸菌DH1株の全ゲノム解析2010

    • Author(s)
      小野直亮, 鈴木真吾, 古澤力, 清水浩, 四方哲也
    • Organizer
      第62回日本生物工学会大会
    • Place of Presentation
      宮崎シーガイアワールドコンベンションセンターサミット
    • Year and Date
      2010-10-28
    • Related Report
      2011 Final Research Report
  • [Presentation] 人工進化実験によって得られたエタノール耐性大腸菌株の網羅的遺伝子発現解析とゲノム異変解析2010

    • Author(s)
      堀之内貴明, 玉岡邦康, 古澤力, 小野直亮, 鈴木真吾, 平沢敬, 四方哲也, 清水浩
    • Organizer
      宮崎シーガイアワールドコンベンションセンターサミット
    • Year and Date
      2010-10-28
    • Related Report
      2011 Final Research Report
  • [Presentation] 人工進化実験によって得られたエタノール耐性大腸菌株の網羅的遺伝子発現解析とゲノム異変解析2010

    • Author(s)
      古澤力
    • Organizer
      第62回日本生物工学会大会
    • Place of Presentation
      宮崎シーガイアワールドコンベンションセンターサミット、宮崎県
    • Year and Date
      2010-10-28
    • Related Report
      2010 Annual Research Report
  • [Presentation] 次世代シーケンサを用いた大腸菌DH1株の全ゲノム解析2010

    • Author(s)
      小野直亮
    • Organizer
      第62回日本生物工学会大会
    • Place of Presentation
      宮崎シーガイアワールドコンベンションセンターサミット、宮崎県
    • Year and Date
      2010-10-28
    • Related Report
      2010 Annual Research Report
  • [Presentation] 複数時間スケールを持つ環境適応ダイナミクス:エタノールストレス環境下での大腸菌人工進化実験における網羅的表現型/遺伝子型解析2010

    • Author(s)
      堀之内貴明, 玉岡邦康, 古澤力, 小野直亮, 鈴木真吾, 平沢敬, 四方哲也, 清水浩
    • Organizer
      第48回生物物理学会年会
    • Place of Presentation
      東北大学
    • Year and Date
      2010-09-20
    • Related Report
      2011 Final Research Report
  • [Presentation] 複数時間スケールを持つ環境適応ダイナミクス:エタノールストレス環境下での大腸菌人工進化実験における網羅的表現型/遺伝子型解析2010

    • Author(s)
      古澤力
    • Organizer
      第48回生物物理学会年会
    • Place of Presentation
      東北大学川内キャンパス、宮城県
    • Year and Date
      2010-09-20
    • Related Report
      2010 Annual Research Report
  • [Presentation] Systems Biotechnology for Creation of Microbial Cell Factroy2010

    • Author(s)
      HiroshiShimizu
    • Organizer
      2010 International Symposium on Advanced Biological Engineering(ISABE' 2010)
    • Place of Presentation
      Beijing, China
    • Year and Date
      2010-07-25
    • Related Report
      2011 Final Research Report
  • [Presentation] Systems Biotechnology for Creation of Microbial Cell Factory2010

    • Author(s)
      清水浩
    • Organizer
      2010 International Symposium on Advanced Biological Engineering (ISABE'2010)
    • Place of Presentation
      Beijing Friendship Hotel, Beijing (China)(招待講演)
    • Year and Date
      2010-07-25
    • Related Report
      2010 Annual Research Report
  • [Presentation] Genome-wide analysis of ethanol stress tolerant strains of Escherichia coli obtained by evolution experiments2010

    • Author(s)
      ChikaraFurusawa
    • Organizer
      Metabolic Engineering VIII
    • Place of Presentation
      Jeju Island, South Korea
    • Year and Date
      2010-06-14
    • Related Report
      2011 Final Research Report
  • [Presentation] Genome-wide analysis of ethanol stress tolerant strains of Escherichia coli obtained by evolution experiments2010

    • Author(s)
      古澤力
    • Organizer
      Metabolic Engineering VIII
    • Place of Presentation
      International Convention Center Jeju, Jeju Island (South Korea)
    • Year and Date
      2010-06-14
    • Related Report
      2010 Annual Research Report
  • [Presentation] 大腸菌エタノールストレス人工進化株の全ゲノム変異解析・遺伝子発現解析2010

    • Author(s)
      堀之内貴明, 玉岡邦康, 古澤力, 平沢敬, 小野直亮, 鈴木真吾, 四方哲也, 清水浩
    • Organizer
      定量生物学の会第二回年会
    • Place of Presentation
      大阪大学
    • Year and Date
      2010-01-09
    • Related Report
      2011 Final Research Report
  • [Presentation] 大腸菌エタノールストレス人工進化株の全ゲノム変異解析・遺伝子発現解析2010

    • Author(s)
      堀之内貴明
    • Organizer
      定量生物学の会 第二回年会
    • Place of Presentation
      大阪大学吹田キャンパス
    • Year and Date
      2010-01-09
    • Related Report
      2009 Annual Research Report
  • [Presentation] Genome-Wide Mutational and Expression Analyses of Evolved Escherichia Coli Strains Under Ethanol Stress2009

    • Author(s)
      Takaaki Horinouchi, Kuniyasu Tamaoka, Chikara Furusawa, Takashi Hirasawa, Naoaki Ono, Shingo Suzuki, Tetsuya Yomo, Hiroshi Shimizu
    • Organizer
      The 20th International Conference on Genome Informatics(GIW 2009)
    • Place of Presentation
      Yokohama, Japan
    • Year and Date
      2009-12-14
    • Related Report
      2011 Final Research Report
  • [Presentation] Genome-wide mutational and expression analyses of evolved Escherichia coli strains under ethanol stress2009

    • Author(s)
      Takaaki Horinouchi
    • Organizer
      The 20th International Conference on Genome Ingormatics(GIW 2009)
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2009-12-14
    • Related Report
      2009 Annual Research Report
  • [Presentation] A model-based analysis method for detection of single-base substitution using resequencing microarrays2009

    • Author(s)
      Naoaki Ono, Shingo Suzuki, Chikara Furusawa, Hiroshi Shimizu, Tetsuya Yomo
    • Organizer
      Asia Pacific Biochemical Engineering Conference 2009(APBioChEC' 09)
    • Place of Presentation
      Kobe, Japan
    • Year and Date
      2009-11-25
    • Related Report
      2011 Final Research Report
  • [Presentation] A model-based analysis method for detection of single-bace substitution using resequencing microarrays2009

    • Author(s)
      Naoaki Ono
    • Organizer
      Asia Pacific Biochemical Engineering Conference 2009(APBioCHEC'09)
    • Place of Presentation
      神戸国際会議場
    • Year and Date
      2009-11-25
    • Related Report
      2009 Annual Research Report
  • [Presentation] 次世代シーケンサにおけるメイトペアの距離情報を利用した配列解析アルゴリズムの開発2009

    • Author(s)
      小野直亮, 鈴木真吾, 古澤力, 清水浩, 四方哲也
    • Organizer
      第47回生物物理学会年会
    • Place of Presentation
      徳島文理大学
    • Year and Date
      2009-11-01
    • Related Report
      2011 Final Research Report 2009 Annual Research Report
  • [Presentation] システムバイオテクノロジーによる生物生産2009

    • Author(s)
      清水浩
    • Organizer
      第61回日本生物工学会大会
    • Place of Presentation
      名古屋大学
    • Year and Date
      2009-09-25
    • Related Report
      2011 Final Research Report
  • [Presentation] システムバイオテクノロジーによる生物生産2009

    • Author(s)
      清水浩
    • Organizer
      日本生物工学会第61回大会
    • Place of Presentation
      名古屋大学
    • Year and Date
      2009-09-25
    • Related Report
      2009 Annual Research Report
  • [Presentation] メイトペア情報に基づいた次世代シーケンサによるresequencing解析アルゴリズムの開発2009

    • Author(s)
      小野直亮, 鈴木真吾, 古澤力, 清水浩, 四方哲也
    • Organizer
      第61回日本生物工学会大会
    • Place of Presentation
      名古屋大学
    • Year and Date
      2009-09-24
    • Related Report
      2011 Final Research Report
  • [Presentation] タイリングアレイを用いたエタノール耐性大腸菌株のゲノム変異解析2009

    • Author(s)
      玉岡邦康, 堀之内貴明, 古澤力, 平沢敬, 小野直亮, 鈴木真吾, 四方哲也, 清水浩
    • Organizer
      第61回日本生物工学会大会
    • Place of Presentation
      名古屋大学
    • Year and Date
      2009-09-24
    • Related Report
      2011 Final Research Report
  • [Presentation] タイリングアレイを用いたエタノール耐性大腸菌株のゲノム変異解析2009

    • Author(s)
      玉岡邦康
    • Organizer
      日本生物工学会第61回大会
    • Place of Presentation
      名古屋大学
    • Year and Date
      2009-09-24
    • Related Report
      2009 Annual Research Report
  • [Presentation] メイトペア情報に基づいてた次世代シーケンサによるresequencing解析アルゴリズムの開発2009

    • Author(s)
      小野直亮
    • Organizer
      日本生物工学会第61回大会
    • Place of Presentation
      名古屋大学
    • Year and Date
      2009-09-24
    • Related Report
      2009 Annual Research Report
  • [Book] 高等研報告書1005,生物進化の持続性と転移,第2章大腸菌の長期植え継ぎ培養における表現型・遺伝子型の網羅的解析2011

    • Author(s)
      古澤力, 堀之内貴明, 清水浩
    • Publisher
      財団法人国際高等研究所
    • URL

      http://www-shimizu.ist.osaka-u.ac.jp/

    • Related Report
      2011 Final Research Report
  • [Book] 高等研報告書1005生物進化の持続性と転移(研究代表者津田一郎)2011

    • Author(s)
      古澤力, 堀之内貴明, 清水浩
    • Publisher
      第2章大腸菌の長期植え継ぎ培養における表現型・遺伝子型の網羅的解析
    • Related Report
      2011 Annual Research Report
  • [Remarks]

    • URL

      http://www-shimizu.ist.osaka-u.ac.jp/

    • Related Report
      2011 Final Research Report
  • [Remarks]

    • URL

      http://www-shimizu.ist.osaka-u.ac.jp/

    • Related Report
      2011 Annual Research Report
  • [Remarks]

    • URL

      http://www-shimizu.ist.osaka-u.ac.jp/

    • Related Report
      2010 Annual Research Report
  • [Remarks]

    • URL

      http://www-shimizu.ist.osaka-u.ac.jp/

    • Related Report
      2009 Annual Research Report

URL: 

Published: 2009-04-01   Modified: 2016-04-21  

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