Development of for creating segmental aneuploid word processing technology of genome in yeast

• Project/Area Number: 24380048
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: Osaka University
• Principal Investigator: HARASHIMA Satoshi 大阪大学, 工学(系)研究科(研究院), 教授 (70116086)
• Co-Investigator(Kenkyū-buntansha): SUGIYAMA Minetaka 大阪大学, 大学院工学研究科, 准教授 (80379130) ; SASANO Yu 大阪大学, 大学院工学研究科, 助教 (90640194) ; KANEKO Yoshinobu 大阪大学, 大学院工学研究科, 寄附講座教授 (90161182) ; MAEKAWA Hiromi 大阪大学, 大学院工学研究科, 寄附講座准教授 (80399683)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2014: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2013: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2012: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
• Keywords: 酵母 / ゲノム工学 / 染色体工学 / 異数体 / バイオテクノロジー / ゲノムの多様性創出工学 / 育種 / 染色体の部分重複

Outline of Final Research Achievements

It is being known that segmental aneuploidy plays an important role for industrial yeast strain to display useful traits. We have developed a news genome engineering technology to create segmental duplication of any desired region of chromosome in yeast. This technology designated PCDup for PCR-mediated chromosome segmental duplication was applied to construct a series of 62 strains harboring every 10 kb to 200 kb chromosomal region. 53 among 62 regions were successfully duplicated while other regions were not. Phenotypic analysis of 53 strains harboring segmental duplication revealed that some of the strains displayed industrially useful trains such as resistance to high temperature and acid environment. Since it has never been reported that over-expression of individual genes present on those regions causes such phenotype, we suggest that those phenotypes were likely due to simultaneous over-expression of some of genes present on those regions.

Research Products

• [Journal Article] Cloning and functional analysis of HpFAD2 and HpFAD3 genes encoding Δ12- and Δ15-fatty acid desaturases in Hansenula polymorpha. (2014)
  • Author(s): Sangwallek, J., Kaneko, Y., Tsukamoto, T., Marui, M., Sugiyama, M., Ono, H., Bamba, T., Fukusaki, E., Harashima, S.
  • Journal Title: Gene Volume: 533 Issue: 1 Pages: 110-118
  • DOI: 10.1016/j.gene.2013.09.115
  • Peer Reviewed
• [Journal Article] Suppression mechanism of the calcium sensitivity in Saccharomyces cerevisiae ptp2Δmsg5Δ double disruptant involves a novel HOG-independent function of Ssk2, transcription factor Msn2 and the protein kinase A component Bcy1 (2014)
  • Author(s): Lavina, W., Shahsavarani, H., Saidi, A., Sugiyama, M., Kaneko, Y., Harashima, S.
  • Journal Title: J. Biosci. Bioeng Volume: 117 Issue: 2 Pages: 135-141
  • DOI: 10.1016/j.jbiosc.2013.06.022
  • NAID: 110009804062
  • Peer Reviewed
• [Journal Article] Nuclear localization of Haa1, which is linked to its phosphorylation status, mediates lactic acid tolerance in Saccharomyces cerevisiae. (2014)
  • Author(s): Sugiyama, M., Akase, SP., Nakanishi, R., Horie, H., Kaneko, Y., Harashima, S.
  • Journal Title: Appl. Environ. Microbiol Volume: 80 Issue: 11 Pages: 3488-3495
  • DOI: 10.1128/aem.04241-13
  • Peer Reviewed
• [Journal Article] Effects of deletion of different PP2C protein phosphatase genes on stress responses in Saccharomyces cerevisiae. (2014)
  • Author(s): Sharmin, D., Sasano, Y., Sugiyama, M., Harashima, S.
  • Journal Title: Yeast Volume: 10 Issue: 10 Pages: 393-409
  • DOI: 10.1002/yea.3032
  • Peer Reviewed
• [Journal Article] Genome-wide mapping of unexplored essential regions in the Saccharomyces cerevisiae genome: evidence for hidden synthetic lethal combinations in a genetic interaction network. (2014)
  • Author(s): Kaboli, S., Yamakawa, T., Sunada, K., Takagaki, T., Sasano, Y., Sugiyama, M., Kaneko, Y., Harashima, S.
  • Journal Title: Nucleic Acids Res Volume: 42 Issue: 15 Pages: 9838-9853
  • DOI: 10.1093/nar/gku576
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] 多様性創出ゲノム工学技術の開発と微生物育種への応用 (2014)
  • Author(s): 笹野 佑、Yeon-Hee Kim、杉山 峰崇、原島 俊
  • Journal Title: 生物工学会誌 Volume: 92(11) Pages: 589-592
  • Peer Reviewed
• [Journal Article] 酵母ゲノム工学の最前線 (2013)
  • Author(s): 杉山 峰崇、笹野 佑、原島 俊
  • Journal Title: 細胞工学 Volume: 32 Pages: 592-599
• [Journal Article] Increased transcription of RPL40A and RPL40B is important for the improvement of RNA production in Saccharomyces cerevisiae. (2013)
  • Author(s): Khatun, F., Kurata, K., Chuwattanakul, V., Sugiyama, M., Kaneko, Y., Harashima, S.
  • Journal Title: J. Biosci. Bioeng. Volume: 116(4) Issue: 4 Pages: 423-432
  • DOI: 10.1016/j.jbiosc.2013.04.006
  • NAID: 110009675454
  • Peer Reviewed
• [Journal Article] Increase in rRNA content in a Saccharomyces cerevisiae suppressor strain from rrn10 disruptant by rDNA cluster duplication. (2013)
  • Author(s): Khatun, F., Sasano, Y., Sugiyama, M., Kaneko, Y., Harashima, S.
  • Journal Title: Appl. Microbiol. Biotechnol. Volume: 97(20) Issue: 20 Pages: 9011-9019
  • DOI: 10.1007/s00253-013-5065-9
  • Peer Reviewed
• [Journal Article] 最新のゲノム編集技術が拓く新たな微生物育種への道 (2013)
  • Author(s): 笹野 佑
  • Journal Title: 生物工学会誌 Volume: 91 Pages: 719-719
• [Journal Article] Large-scale genome reorganization in Saccharomyces cerevisiae through combinatorial loss of mini-chromosomes (2012)
  • Author(s): Youji Ueda, Shigehito Ikushima, Minetaka Sugiyama, Ryo Matoba, Yoshinobu Kaneko, Kenichi Matsubara, Satoshi Harashima
  • Journal Title: Journal of Bioscience and Bioengineering Volume: VOL. 113 Issue: 6 Pages: 675-682
  • DOI: 10.1016/j.jbiosc.2012.01.013
  • NAID: 10031085641
  • Peer Reviewed
• [Journal Article] Creation af an ethariol-tolerant yeast strain by genome reconstruction based on chromosome splitting technology. (2012)
  • Author(s): Park, AH., Sugiyama, M , Harashima, S., Kim, YH.
  • Journal Title: J. Microbiol. Biotechnol. Volume: 22(2) Pages: 184-9
  • Peer Reviewed
• [Presentation] CRISPR-PCS法による酵母染色体複数部位の同時分断 (2014)
  • Author(s): 笹野 佑、長澤宏器、Saeed Kaboli、杉山峰崇、原島 俊
  • Organizer: 日本生物工学会第66回大会
  • Place of Presentation: 札幌コンベンションセンター（札幌市）
  • Year and Date: 2014-09-09 – 2014-09-11
• [Presentation] ゲノムエンジニアリングによる出芽酵母二倍体ゲノムの部分一倍体化と育種への応用 (2014)
  • Author(s): 砂田啓輔、Saeed Kaboli、宮本哲也、笹野 佑、杉山峰崇、原島 俊
  • Organizer: 日本生物工学会第66回大会
  • Place of Presentation: 札幌コンベンションセンター（札幌市）
  • Year and Date: 2014-09-09 – 2014-09-11
• [Presentation] ゲノムシャフリングによる出芽酵母の高温耐性の改良と高温アルコール発酵 (2014)
  • Author(s): 杉山峰崇、呉 俊元、Badua Mark Anthony A.、笹野 佑、金子嘉信、原島 俊
  • Organizer: 日本生物工学会第66回大会
  • Place of Presentation: 札幌コンベンションセンター（札幌市）
  • Year and Date: 2014-09-09 – 2014-09-11
• [Presentation] 高温耐性出芽酵母C3723株におけるCDC19-C遺伝子による高温性獲得機構 (2014)
  • Author(s): Wang Kun、木村駿太、笹野 佑、杉山峰崇、原島 俊
  • Organizer: 日本生物工学会第66回大会
  • Place of Presentation: 札幌コンベンションセンター（札幌市）
  • Year and Date: 2014-09-09 – 2014-09-11
• [Presentation] 酵母を用いた真核生物遺伝子機能の解明とバイオテクノロジーへの応用 (2014)
  • Author(s): 原島 俊
  • Organizer: 日本生物工学会第66回大会
  • Place of Presentation: 札幌コンベンションセンター（札幌市）
  • Year and Date: 2014-09-09 – 2014-09-11
  • Invited
• [Presentation] 酵母バイオテクノロジーの発展とともに40年 (2014)
  • Author(s): 原島 俊
  • Organizer: 第21回酵母合同シンポジウム
  • Place of Presentation: 東京大学 伊藤国際学術研究センター 伊藤謝恩ホール（東京都）
  • Year and Date: 2014-09-03 – 2014-09-04
  • Invited
• [Presentation] ゲノム工学技術を用いた出芽酵母新規合成致死遺伝子の探索 (2014)
  • Author(s): 高垣太緒、Saeed Kaboli、砂田啓輔、山川卓也、笹野 佑、杉山峰崇、原島 俊
  • Organizer: 酵母遺伝学フォーラム第47回研究報告会
  • Place of Presentation: 東京大学弥生会館（東京都）
  • Year and Date: 2014-09-01 – 2014-09-02
• [Presentation] Segmental haploidization of diploid genome in Saccharomyces cerevisiae and its application to breeding (2014)
  • Author(s): Saeed Kaboli、Tetsuya Miyamoto、Keisuke Sunada、Yu Sasano、Minetaka Sugiyama、Satoshi Harashima
  • Organizer: 酵母遺伝学フォーラム第47回研究報告会
  • Place of Presentation: 東京大学弥生会館（東京都）
  • Year and Date: 2014-09-01 – 2014-09-02
• [Presentation] 出芽酵母におけるミニ染色体のワンステップ作成技術の開発 (2014)
  • Author(s): 長澤宏器、笹野 佑、Saeed Kaboli、杉山峰崇、原島 俊
  • Organizer: 酵母遺伝学フォーラム第47回研究報告会
  • Place of Presentation: 東京大学弥生会館（東京都）
  • Year and Date: 2014-09-01 – 2014-09-02
• [Presentation] 過剰発現で乳酸耐性を付与する出芽酵母転写活性化因子Haa1の機能解析 (2014)
  • Author(s): 中西良太、堀江仁志、松田祐典、笹野 佑、杉山峰崇、原島 俊
  • Organizer: 酵母遺伝学フォーラム第47回研究報告会
  • Place of Presentation: 東京大学弥生会館（東京都）
  • Year and Date: 2014-09-01 – 2014-09-02
• [Presentation] 高温耐性出芽酵母C3723株におけるCDC19-C遺伝子による高温耐性化 (2014)
  • Author(s): 木村駿太、王 琨、○笹野 佑、杉山峰崇、原島 俊
  • Organizer: 酵母遺伝学フォーラム第47回研究報告会
  • Place of Presentation: 東京大学弥生会館（東京都）
  • Year and Date: 2014-09-01 – 2014-09-02
• [Presentation] ゲノムシャフリングによる出芽酵母高温耐性の改良と高温アルコール発酵 (2014)
  • Author(s): 杉山峰崇、呉 俊元、Badua Mark Anthony A.、中西良太、笹野 佑、金子嘉信、原島 俊
  • Organizer: 酵母遺伝学フォーラム第47回研究報告会
  • Place of Presentation: 東京大学弥生会館（東京都）
  • Year and Date: 2014-09-01 – 2014-09-02
• [Presentation] Recent advances in breeding technologies for microorganisms More than just a trend- (2013)
  • Author(s): Satoshi Harashima
  • Organizer: JSPS-NRCT Young Scientist Seminar 2013
  • Place of Presentation: 大阪
• [Presentation] Genome-wide construction of a series of segmental aneuploids for genome analysis and breeding in Saccharomyces cerevisiae. (2013)
  • Author(s): Waranya Natesuntorn, Kotaro Iwami, Yuki Matsubara, Tetsuya Hayashi, Yu Sasano, Minetaka Sugiyama, Yoshinobu Kaneko, Satoshi Harashima
  • Organizer: JSPS-NRCT Young Scientist Seminar 2013
  • Place of Presentation: 大阪
• [Presentation] Genome-wide mapping for unexplored essential regions harboring synthetic lethal interactions in Saccharomyces cerevisiae. (2013)
  • Author(s): Saeed Kaboli, Takuya Yamakawa, Deasty Imara, Yu Sasano, Minetaka Sugiyama, Yoshinobu Kaneko, Satoshi Harashima
  • Organizer: JSPS-NRCT Young Scientist Seminar 2013
  • Place of Presentation: 大阪
• [Presentation] Molecular breeding of Saccharomyces cerevisiae strain producing high level of RNA. (2013)
  • Author(s): Yu Sasano, Fahmida Khatun, Shogo Usugi, Minetaka Sugiyama, Satoshi Harashima
  • Organizer: 26th International Conference on Yeast Genetics and Molecular Biology
  • Place of Presentation: Frankfurt, Germany
• [Presentation] PCR-mediated chromosome duplication method generates a defined duplicated chromosome as a novel tool for breeding and genome analysis in Saccharomyces cerevisiae. (2013)
  • Author(s): Waranya Natesuntorn, Kotaro Iwami, Yuki Matsubara, Yu Sasano, Minetaka Sugiyama, Yoshinobu Kaneko, Satoshi Harashima
  • Organizer: 26th International Conference on Yeast Genetics and Molecular Biology
  • Place of Presentation: Frankfurt, Germany
• [Presentation] CRISPR/Cas-PCSシステムを利用した染色体複数部位同時分断技術の開発 (2013)
  • Author(s): 笹野 佑、長澤宏器、杉山峰崇、原島 俊
  • Organizer: 酵母遺伝学フォーラム第46回研究報告会
  • Place of Presentation: 仙台
• [Presentation] Novel landscape for genetic interactions in Saccharomyces cerevisiae. (2013)
  • Author(s): Saeed Kaboli, Takuya Yamakawa, Deasty Imara, Yu Sasano, Minetaka Sugiyama, Yoshinobu Kaneko, Satoshi Harashima
  • Organizer: YEAST GENETICS AND MOLECULAR BIOLOGY NEWS JAPAN No.46
  • Place of Presentation: 仙台
• [Presentation] Genome-wide construction of a series of segmental aneuploids in Saccharomyces cerevisiae. (2013)
  • Author(s): Waranya Natesuntorn, Yuki Matsubara, Yu Sasano, Minetaka Sugiyama, Yoshinobu Kaneko, Satoshi Harashima
  • Organizer: YEAST GENETICS AND MOLECULAR BIOLOGY NEWS JAPAN No.46
  • Place of Presentation: 仙台
• [Presentation] Systematic segmental duplication of chromosomes for genome analysis and breeding in Saccharomyces cerevisiae (2013)
  • Author(s): Waranya Natesuntorn, Yuki Matsubara, Yu Sasano, Minetaka Sugiyama, Satoshi Harashima
  • Organizer: 日本生物工学会第65回大会
  • Place of Presentation: 広島
• [Presentation] 酵母rrn10遺伝子破壊抑圧変異株におけるrDNA領域の重複によるRNA量の増加 (2013)
  • Author(s): Khatun Fahmida、笹野 佑、杉山峰崇、原島 俊
  • Organizer: 日本生物工学会第65回大会
  • Place of Presentation: 広島
• [Presentation] rDNA写活必須遺伝子RRN5の破壊抑圧変異株を利用したRNA高含有出芽酵母の分子育種 (2013)
  • Author(s): 宇杉祥吾、Khatun Fahmida、狩屋貴洋、笹野 佑、杉山峰崇、原島 俊
  • Organizer: 日本生物工学会第65回大会
  • Place of Presentation: 広島
• [Presentation] 出芽酵母におけるゲノム網羅的な染色体部分重複と表現型解析 (2013)
  • Author(s): 杉山峰崇、Waranya Natesuntorn、松原裕樹、林 達也、原島 俊
  • Organizer: 日本生物工学会第65回大会
  • Place of Presentation: 広島
• [Presentation] 出芽酵母におけるゲノム網羅的な染色体部分重複と表現型解析 (2013)
  • Author(s): 杉山峰崇、Waranya Natesuntorn、松原裕樹、林 達也、原島 俊
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸
• [Presentation] Random Walks on Chromosomal Regions Reveal Hidden Lethal Relationships among Non-essential Genes in the Genome of Saccharomyces cerevisiae (2013)
  • Author(s): Saeed Kaboli, Takuya Yamakawa, Minetaka Sugiyama, Yoshinobu Kaneko, Satoshi Harashima
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸
• [Presentation] 酵母の育種理論と育種技術の発展とともに35年 (2012)
  • Author(s): 原鳥俊
  • Organizer: 第19回日本生物工学会九州支部福岡大会特別講演
  • Place of Presentation: 別府大学(招待講演)
  • Year and Date: 2012-12-01
• [Presentation] ウイルス学におけるdual use 「ゲノムの多様性創出工学と生命科学・生命工学におけるデュアルユース問題」 (2012)
  • Author(s): 原島俊
  • Organizer: 第60回日本ウィルス学会学術集会特別シンポジウム
  • Place of Presentation: グランキューブ大阪(招待講演)
  • Year and Date: 2012-11-14
• [Presentation] Exploitation of PCDup technology for breeding and genome analysis in Saccharonyces cerevisiae (2012)
  • Author(s): W. Natesuntorn. Y. Matsubara, T. Hayasbi. M. Sugiyama, Y. Kaleko, S. Harashima
  • Organizer: 日本生物工学会第64回大会
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2012-10-25
• [Presentation] Exploitation of PCDup technology for breeding and genome analysis in Saccharonyces cerevisiae. (2012)
  • Author(s): W. Natesuatorn, Y. Matsubara, T. Hayashi, M. Sugiyama, Y. Kaneko, S. Harashima
  • Organizer: YEAST GENETICS AND MOLECULAR BIOLOGY NEWS JAPAN No. 45
  • Place of Presentation: 京都大学宇治キャンパス
  • Year and Date: 2012-09-06
• [Presentation] Recent Advances on New Breeding Technologies in Microorganisns -More than lust a Trend- (2012)
  • Author(s): Satoshi Harashima
  • Organizer: International Symposium & Annual Meeting A New Era of Biotechnology and Bioeconomy
  • Place of Presentation: Busan, Korea(招待講演)
  • Year and Date: 2012-06-28
• [Presentation] 酵母におけるゲノム工学技術の開発とゲノム機能の解明・育種への応用
  • Author(s): 原島 俊
  • Organizer: 大阪大学大学院工学研究科 核酸制御(陽進堂)共同研究講座設立記念シンポジウム
  • Place of Presentation: 大阪
• [Presentation] 多様性創出ゲノム工学技術の開発と微生物育種への応用
  • Author(s): 原島 俊
  • Organizer: 2013年度JBA新資源生物変換研究会シンポジウム
  • Place of Presentation: 東京
• [Book] Microbial production -From genome design to cell engineering-, Microbial Production (2014)
  • Author(s): Sasano Y, Sugiyama M. Harashima S.
  • Total Pages: 550
  • Publisher: Springer
• [Book] 微生物を活用した新世代の有用物質生産技術 (2012)
  • Author(s): 原島俊・Yeon-Hee Kim・西沢正文
  • Publisher: (株)シーエムシー出版
• [Book] 合成生物工学の隆起-有用物質の新たな生産法構築をめざして- (2012)
  • Author(s): 原島俊・西沢正文
  • Publisher: (株)シーエムシー出版
• [Remarks] 大阪大学工学研究科ゲノム機能工学研究室（原島研究室）
  • URL: http://www.bio.eng.osaka-u.ac.jp/mg/
• [Remarks] 大阪大学・原島研究室
  • URL: http://www.bio.eng.osaka-u.ac.jp/mg/