| Project/Area Number |
12460044
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | Osaka University |
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Principal Investigator |
HARASHIMA Satoshi Osaka University, Department of Biotechnology, Professor, 大学院・工学研究科, 教授 (70116086)
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| Co-Investigator(Kenkyū-buntansha) |
NISHIZAWA Masafumi Keio University School of Medicine, Department of Microbiology and Immunology, Lecturer, 医学部・微生物学教室, 講師 (20218150)
KANEKO Yoshinobu Osaka University, Department of Biotechnology, Associate Professor, 大学院・工学研究科, 助教授 (90161182)
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥14,100,000 (Direct Cost: ¥14,100,000)
Fiscal Year 2002: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2001: ¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 2000: ¥6,100,000 (Direct Cost: ¥6,100,000)
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| Keywords | yeast / chromosome-splitting / minimal genome / chromosome shuffling / 遺伝子破壊 / 染色体の分断 |
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| Research Abstract |
Chromosome engineering techniques that can manipulate a large segment of chromosomal DNA are useful not only for studying the organization of eukaryotic genomes but also for the improvement of industrially important strains. Toward the development of techniques that can efficiently manipulate a large segment of chromosome, we have developed in this study a one-step chromosome splitting technique in a haploid Saccharomyces cerevisiae. With this technique we could successfully to create a novel yeast haploid strain having up to 21 chromosomes. To facilitate chromosome splitting at multiple sites on chromosomes, we further focused on the δ sequences that are distributed in more than 200 copies throughout the yeast genome. We constructed a new chromosome-splitting vector harboring the δ sequence and the reusable marker cassette for transformation, and successfully split several chromosomes. We have also successfully developed a novel technique for shuffling a desired chromosomal region of two yeast strains. Chromosome shuffling technique provide a powerful tool to evaluate the consequence of replacement of a particular chromosomal region in yeast. Although achievement of this study is still far from ultimate aim, i.e., creation of minimal genome yeast and exploration of relationship between genome structure and function in eukaryote, we believe that fundamental technology for engineering genome toward the goal was established through this study.
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