• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Creation of minimal genome yeast by chromosome splitting technology

Research Project

Project/Area Number 12460044
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionOsaka University

Principal Investigator

HARASHIMA Satoshi  Osaka University, Department of Biotechnology, Professor, 大学院・工学研究科, 教授 (70116086)

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)
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
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)
Keywordsyeast / chromosome-splitting / minimal genome / chromosome shuffling / 遺伝子破壊 / 染色体の分断
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.

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (47 results)

All Other

All Publications (47 results)

  • [Publications] Mizuno, T., Harashima, S.: "Activation of basal transcription by a mutation in SIN4, a yeast global repressor, occurs through a mechanism different from activator-mediated transcriptional enhancement"Mol.Gen.Genet.. 263. 48-59 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kunoh, T., Kaneko, Y., Harashima, S.: "YHP1 encodes a new homeopraotein that binds to the IME1 promoter in saccharomyces cerevisiae"Yeast. 16. 439-449 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kunoh, T., Sakuno, T., Furukawa, T., Kaneko, Y., Harashima, S.: "Genetic characterization of rbt mutants that enhance basal transcription from core promoters in Saccharomyces cerevisiae"J.Biochem.. 128. 575-584 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tamai, Y., Tanaka, K., Umemoto, N., Tomizuka, K., Kaneko, Y.: "Diversity of the HO gene encoding an endonuclease for mating-type conversion in the bottom fermenting yeast Saccharomyces pastorianus"Yeast. 16. 1335-1343 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Lu, S.-F., Tolstorukov, I.I., Anamnart, S., Kaneko, Y., Harashima, S.: "Cloning sequencing and functional analysis of H-OLE1 gene encoding Δ9-fatty acid desaturase in Hansenula polymorpha"Appl.Microbiol.Biotechnol.. 54. 499-509 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakagawa, Y., Sugioka, S., Kaneko, Y., Harashima, S.: "O2R, a novel regulatory element mediating Rox1p-Independent O2 and unsaturated fatty acid repression of OLE1 in Saccharomyces cerevisiae"J.Bacteriol.. 183. 745-751 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Harashima, S., Kaneko, Y.: "Application of the PHO5-gene-fusion technology to molecular genetics and biotechnology of yeast"J.Biosci.Bioeng.. 91. 325-338 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tamai Y, Tanaka K, Kaneko Y, Harashima S.: "HO gene polymorphism in Saccharomyces industrial yeasts and applicalion of novel HO genes to convert homothallism to heterothallism in combination with the mating-type detection cassette"Appl.Microbiol.Biotechnol.. 55. 333-340 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakumoto N., Yamashita, H., Mukai, Y., Kaneko, Y., Harashima, S.: "Dual-specificity protein phosphatase Yvh1p, which is required for vegetative growth and sporulation, interacts with yeast pescadillo homolog in Saccharomyces cerevisiae"Biochem.Biophys.Res.Commun.. 289. 608-615 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Shibasaki S, Ninomiya Y, Ueda M, Iwahashi M, Katsuragi T, Tani Y, Harashima S, Tanaka A.: "Intelligent yeast strains with the ability to self-monitor the concetrations of intra-and extracellular phosphate or ammonium ion by emission of fluorescence from the cell surface"Appl.Microbiol.Biotechnol.. 57. 702-707 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakagawa, Y., Sakumoto, N., kaneko, Y., Harashima, S.: "Mga2p is a putative sensor for low temperature and oxygen to induce OLE1 transcription in Saccharomyces cerevisiae"Biochem.Biophys.Res.Commun.. 291. 707-713 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakagawa Y.et al.: "Mga2p Is a Putative Sensor for Low Temperature and Oxygen to Induce OLE1 Transcription in Saccharomyces cerevisiae"Biochem.Biophys.Res.Commun.. 291. 707-713 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakumoto N.et al.: "A series of double disruptants for protein phosphatase genes in Saccharomyces cerevisiae and their phenotypic analysi"Yeast. 19. 587-599 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Saskiawan I.et al.: "Chemo-enzymatic synthesis of the glycosylated α-mating factor of Saccharomyces cerevisiae and analysis of its biological activity"Arch.Biochem.Biophys. 406. 127-134 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Widianto D.et al.: "Creating a Saccharomyces cerevisiae Haploid Strain Having 21 Chromosomes"J.Biosci.Bioeng.. 95. 1-6 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Mizuno T, Harashima S.: "Ga111 is a general activator for basal transcription whose activity is regulated by a general repressor Sin4 in yeast"Molecular and General Genomics. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Mizuno, T., Harashima, S.: "Activation of basal transcription by a mutation in SIN4, a yeast global repressor, occurs through a mechanism different from activator-mediated transcriptional enhaucement"Mol. Gen. Genet.. 263. 48-59 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kunoh T., Kaneko, Y., Harashima, S.: "YHP1 encodes a new homeoprotein that binds to the IME1 promoter in Saccharomyces cerevisiae"Yeast. 16. 439-449 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kunoh, T., Sakuno, T., Furukawa, T., Kaneko, Y., Harashima, S.: "Genetic Characterization of rbt mutants that enhance basal transcription from core promoters in Saccharomyces cerevsiae"J. Biochem. 128. 575-584 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tamai, Y., Tanaka, K., Umemoto, N., Tomizuka, K., Kaneko, Y.: "Diversity of the HO gene encoding, an endonuclease for mating-type conversion in the bottom fermenting yeast Saccharomyces pastorianus"Yeast. 16. 1335-1343 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Lu, S.- F., Tolstorukov, I. I., Anamnart, S., Kaneko, Y., Harashima, S.: "Cloning, sequencing, and functional analysis of H-OLE1 gene encoding Δ9-fatty acid desaturase in Hansenula polymorpha"Appl. Microbiol. Biotechnol.. 54. 499-509 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kaneko, Y., Kobayashi, H., Kiatpapan, P., Nishimoto, T., Napitupulu, R., Ono, H., Murooka, Y.: "Development of a host-vector system for Lactobacillus plantarum L137 isolated from a traditional fermented food produced in the Philippines"J. Biosci. Bioeng.. 89. 62-67 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakagawa, Y., Sugioka, S., Kaneko, Y., Harashima, S.: "O2R. a novel regulatory element mediating Roxlp-Independent O2 and unsaturated fatty acid repression of OLE1 in Saccharomyces cerevisiae"J. Bacteriol. 183. 745-751 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Harashima, S., Kaneko, Y.: "Application of the PHO5-gene-fusion technology to molecular genetics and biotechnology of yeast"J. Biosci. Bioeng.. 91. 325-338 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tamai Y, Tanaka K, Kaneko Y, Harashima S.: "HO gene polymorphism in Saccharomyces industrial yeasts and application of novel HO genes to convert homothallism to heterothallism in combination with the mating-type detection cassette"Appl. Microbiol. Biotechnol.. 55. 333-340 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakumoto N., Yamashita, H., Mukai, Y., Kaneko, Y., Harashima, S.: "Dual-specificity protein phosphatase Yvhlp, which is required for vegetative growth and sporulation, interacts with yeast pescadillo homolog in Saccharomyces cerevisiae"Biochem. Biophys. Res. Commun.. 289. 608-615 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Shibasaki S, Ninomiya Y, Ueda M, Iwahashi M, Katsuragi T, Tani Y, Harashima S, Tanaka A.: "Intelligent yeast strains with the ability to self-monitor the concentrations of intra- and extracellular phosphate or ammonium ion by emission of fluorescence from the cell surface"Appl. Microbiol. Biotechnol.. 57. 702-707 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakagawa, Y., Sakumoto, N., Kaneko, Y., Harashima, S.: "Mga2p is a putative sensor for low temperature and oxygen to induce OLE1 transcription in Saccharomyces cerevisiae"Biochem. Biophys. Res. Commun.. 291. 707-713 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakagawa Y. et al,: "Mga2p Is a Putative Sensor for Low Temperature and Oxygen to Induce OLE1 Transcription in Saccharomyces cerevisiae"Biochem. Biophys. Res. Commun.. 291. 707-713 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakumoto N.et al,: "A series of double disruptants for protein phosphatase genes in Saccharomyces cerevisiae and their phenotypic analysis"Yeast. 19. 587-599 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Saskiawan I. et al.: "Chemo-enzymatic synthesis of the glycosylated α-mating factor of Saccharomyces cerevisiae and analysis of its biological activity"Arch. Biochem. Biophys.. 406. 127-134 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Widianto D. et al,: "Creating a Saccharomyces cerevisiae Haploid Strain Having 21 Chromosomes"J. Biosci. Bioeng.. 95. 1-6 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Mizuno T & Harashima S.: "Gal11 is a general activator for basal transcription whose activity is regulated by a general repressor Sin4 in yeast"Molecular and General Genomics. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakagawa Y. et al.: "Mga2p Is a Putative Sensor for Low Temperature and Oxygen to Induce OLE1 Transcription in Saccharomyces cerevisiae"Biochem.Biophys.Res.Commun.. 291. 707-713 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sakumoto N. et al.: "A series of double disruptants for protein phosphatase genes in Saccharomyces cerevisiae and their phenotypic analysis"Yeast. 19. 587-599 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Saskiawan I. et al.: "Chemo-enzymatic synthesis of the glycosylated α-mating factor of Saccharomyces cerevisiae and analysis of its biological activity"Arch.Biochem.Biophys.. 406. 127-134 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Widianto D. et al.: "Creating a Saccharomyces cerevisiae Haploid Strain Having 21 Chromosomes"J.Biosci.Bioeng.. 95. 1-6 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Mizuno T., Harashima S.: "Gal11 is a general activator for basal transcription whose activity is regulated by a general repressor Sin4 in yeast"Molecular and General Genomics. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nakagawa, Y. et al.: "O2R a regulatory element mediating Rox-lp-independent O2 and unsaturated fatty acid repressions of OLEl, in Saccharomyces cerevisiae"J.Bacrteriol.. 183. 745-751 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Harashima, S., Kaneko Y.: "Application of the PHO5-gene-fusion technology to molecular genetics and biotechnology of yeast"J.Biosci.Bioeng.. 91. 325-338 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tamai Y, et al.: "HO gene polymorphism in Saccharomyces industrial yeasts and application of novel HO genes to convert homothallism to heterothallism in combination with the mating-type detcction cassette"Appl Microbiol Biotechnol.. 55. 333-340 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Sakumoto N. et al.: "Dual-specificity protein phosphatase Yvhlp, which is required for vegetative growth and sporulation, interacts with yeast pescadillo homolog in Saccharomyces cerevisiae"Biochem Biophys Res Commun.. 289. 608-615 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Shibasaki S, et al.: "Intelligent yeast strains with the ability to self-monitor the concentrations of intra-and extracellular phosphate or ammonium ion by emission of fluorescence from the cell surface"Appl Microbiol Biotechnol.. 57. 702-707 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kunoh,I.,Kaneko,Y.,Harashima,S.: "YHP1 encodes a new homeoprotein that binds to the IME1 promoter in Saccharomyces cerevisiae."Yeast. 16:. 439-449 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Mizuno,T.,Harashima,S.: "Activation of basal transcription by a mutation in SIN4, a yeast global repressor, occurs through a mechanism different from activator-mediated transcriptional"Molec.Gen.Genet.. 263. 48-59 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kunoh,I.,Kaneko,Y.,Harashima,S.: "Genetic characterization of rbt mutants that enhance basal transcription from core promoters in Saccharomyces cerevisiae."J.Biochem.. 128(4). 575-584 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Nakagawa,Y.,Sugioka S.,Kaneko,Y.,Harashima,: "O2R, a regulatory element mediating Rox-1p-independent O2 and unsaturated fatty acid repressions of OLE1, in Saccharomyces cerevisiae."J.Bacrteriol.. 183. 745-751 (2001)

    • Related Report
      2000 Annual Research Report

URL: 

Published: 2000-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi