| Project/Area Number |
25250028
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
System genome science
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| Research Institution | Nara Institute of Science and Technology |
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Principal Investigator |
Mori Hirotada 奈良先端科学技術大学院大学, バイオサイエンス研究科, 教授 (90182203)
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| Co-Investigator(Kenkyū-buntansha) |
YAMADA Mamoru 山口大学, 医学(系)研究科, 教授 (30174741)
MATSUNO Hiroshi 山口大学, 理工学研究科, 教授 (10181744)
TAMURA Takeyuki 京都大学, 化学研究所, 助教 (00437261)
TOHSATO Yukari 理化学研究所, 生命システム研究センター, 研究員 (80346171)
MAKI Yasushi 大阪医科, 大学物理学教室, 講師 (60401733)
NAKAHIGASHI Kenji 慶應義塾大学, 先端生命研, 准教授 (70322740)
SAITO Natsumi 鶴岡高専天, 准教授 (50287546)
片岡 正和 信州大学, 工学部, 准教授 (90332676)
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| Co-Investigator(Renkei-kenkyūsha) |
KOSAKA Tomouki 山口大学, 農学部, 助教 (70500453)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥46,410,000 (Direct Cost: ¥35,700,000、Indirect Cost: ¥10,710,000)
Fiscal Year 2015: ¥13,650,000 (Direct Cost: ¥10,500,000、Indirect Cost: ¥3,150,000)
Fiscal Year 2014: ¥14,430,000 (Direct Cost: ¥11,100,000、Indirect Cost: ¥3,330,000)
Fiscal Year 2013: ¥18,330,000 (Direct Cost: ¥14,100,000、Indirect Cost: ¥4,230,000)
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| Keywords | E. coli / bar-code / deletion collection / population dynamics / chemical genomics / stationary phase / deep sequencing / バーコード解析 / 大腸菌population変動解析 / 長期定常期 / ケミカルバイオロジー / 大腸菌 / 分子バーコード / 一遺伝子欠失株 / deep sequence / 貞コピー可動型発現プラスミド / population変動 / Multiplex解析 / Chemical Genomics / 二重欠失株 / 遺伝的ネットワーク / 細胞内ネットワーク |
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| Outline of Final Research Achievements |
Comprehensive single gene knockout strain collection with 20nt molecular bar-code of E. coli was constructed. The 20nt random bar-code sequences were determined after validation of the target gene deletion by genomic PCR. About 3600 genes out of 4100 ORF genes were established as knockout strain with bar-code. To prove the value of this collection, we performed analysis 1) to monitor population dynamics during the long term stationary phase in LB up to three weeks, 2) profile analysis in medium containing sub-lethal level of toxic compound. We also determine the host strain genome to confirm its genetic background. For the analysis of the population dynamics in the long term stationary phase, at each of samplings time, half of each of the samples were diluted with the fresh LB medium and re-grow up to OD600 = 2.0. Then re-grown samples were performed bar-seq analysis to quantify the growth ability of each of deletion strains after the passage of stationary phase.
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