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IDENTIFICATION OF DISEASE GENE USING BAC TRANSGENIC MICE

Research Project

Project/Area Number 11694296
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory animal science
Research InstitutionKUMAMOTO UNIVERSITY

Principal Investigator

YAMAMURA Kenichi  KUMAMOTO UNIVERSITY, INSITUTE OF MOLECULAR EMBRYOLOGY AND GENETICS, PROFESSOR, 発生医学研究センター, 教授 (90115197)

Co-Investigator(Kenkyū-buntansha) OHBO Kazuyuki  KUMAMOTO UNIVERSITY, INSTITUTE OF MOLECULARA EMBRYOLOGY AND GENGETICS, ASSITATNT PROFESSOR, 発生医学研究センター, 助手 (70250751)
ARAKI Kimi  KUMAMOTO UNIVERSITY, INSTITUTE OF MOLECULARA EMBRYOLOGY AND GENGETICS, ASSISTANT PROFESSOR, 発生医学研究センター, 助手 (90211705)
ABE Kuniya  KUMAMOTO UNIVERSITY, INSTITUTE OF MOLECULARA EMBRYOLOGY AND GENGETICS, ASSOCIATE PROFESSOR, 発生医学研究センター, 助教授 (40240915)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 2000: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1999: ¥3,300,000 (Direct Cost: ¥3,300,000)
KeywordsBAC / YAC / TRANSGENIC MOUSE / OUAKING / T COMPLEX / MICROINJECTION / ミエリン / tail kinks / Cre-loxP / BAC / 変異マウス / 発生 / ポジショナルクローニング
Research Abstract

Chromosome should be replicated without any loss of DNA at the end of chromosome and each one chromosome should be separated into daughter cells while cell division. To accomplish this process, at least three elements, replication origin, telomere, and centromere, are required. In yeast and E.coli, these elements are well characterized and are already cloned. Using these elements, it is now possible to construct artificial chromosome such as yeast artificial chromosome (YAC) or bacterial artificial chromosome (BAC). These artificial chromosomes can be used for isolating a large fragment of DNA spanning to a few hundreds of kb in BAC and a few magabase in YAC.As YAC is quite unstable, use of BAC is becoming more popular. We have tried to establish a method to produce transgenic mice by microinjecting BAC into fertilized eggs. We found that isolation of BAC can be done by pulse field gel electrophoresis, removal of agarose gel fraction containing BAC with about 100 to 200 kb DNA, and digestion of agarose with agarase. The optimum concentration of BAC DNA for microinjection was 1 to 1.5 ng/μl. Although the number of live born mice is lower than that in injection with normal DNA, the overall efficiency of transgenic production was the same and 10 to 60 % of live mice were transgenic. As BAC9 of about 165kb size seems to contain whole qkI gene, we injected these BAC9 and established two lines of transgenic mice. These mice were mated with quaking mice to produce transgenic mice with quaking background. These transgenic mice did not develop quaking phenotype. BAC containing T gene could also rescue the T phenotype. These results clearly suggest that BAC transgenesis can be a powerful tool to test whether the BAC used for transgenesis contains a candidate gene.

Report

(3 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Kondo T. et al.: "Genomic organization and expression analysis of the mouse qkI locus."Mammal.Genome. 10. 662-669 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Oike,Y. et al.: "Truncated CBP protein leads to classical Rubinstein-Taybi syndrome phenotypes in mice : Implication of a dominant negative mechanism."8. 387-396 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Shimada,H. et al.: "Comparison of ES cell fate in sandwiched aggregates and co-cultured aggregates during blastocyst formation by monitored GFP expression."Mol.Reprod.Dev.. 52. 376-382 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Oike,Y. et al.: "Mice homozygous for a truncated form of a CREB-binding protein (CBP) exhibit defects in hematopoiesis and vasculo-angiogenesis."Blood. 14. 2771-2779 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yamamura,K.: "Overview of trangenic and gene knockout mice."Prog.Exp.Tumor Res.. 35. 13-24 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yamauchi,Y. et al.: "A novel transgenic technique that allows specific marking of the neural crest cell lineage in mice."Dev.Biol.. 212. 191-203 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Araki,K. et al.: "Exchangeable gene trap using the Cre/mutated lox system."Cell.Mol.Biol.. 45. 737-750 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Imaizumi,T., et al.: "Mutant mice lacking Crk-II caused by the gene trap insertional mutagenesis : crk-II is not essential for embryonic development."Biochem.Biophysic.Res.Comm.. 266. 569-574 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hisahara,S. et al.: "Targeted expression of baculovirus p35 caspase inhibitor in oligodendrocytes protects mice against autoimmune-mediated demyelinaion."EMBO J.. 19. 341-348 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] McIlroy,D. et al.: "An auxiliary mode of apoptotic DNA fragmentation provided by phagocytes. :"Genes Dev.. 14. 549-558 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Abe,K. et al.: "Molecular and embryological characterization of a new transgene-induced null allele of mouse Brachyury locus."Mammalian Genome. 11. 238-240 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kondo T.et al.: "Genomic organization and expression analysis of the mouse qkI locus."Mammal.Genome. 10. 662-669 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Oike, Y.et al.: "Truncated CBP protein leads to classical Rubinstein-Taybi syndrome phenotypes in mice : Implication of a dominant negative mechanism."Human Mol.Genet.. 8. 387-396 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Shimada, H.et al.: "Comparison of ES cell fate in sandwiched aggregates and co-cultured aggregates during blastocyst formation by monitored GFP expression."Mol.Reprod.Dev.. 52. 376-382 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Oike, Y.et al.: "Mice homozygous for a truncated form of a CREB-binding protein (CBP) exhibit defects in hematopoiesis and vasculo-angiogenesis."Blood. 93. 2771-2779 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yamamura, K.: "Overview of trangenic and gene knockout mice."Prog.Exp.Tumor Res.. 35. 13-24 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yamauchi, Y.et al.: "A novel transgenic technique that allows specific marking of the neural crest cell lineage in mice."Dev.Biol.. 212. 191-203 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Araki, K.et al.: "Exchangeable gene trap using the Cre/mutated lox system."Cell.Mol.Biol.. 45. 737-750 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Imaizumi, T., et al.: "Mutant mice lacking Crk-II caused by the gene trap insertional mutagenesis : crk-II is not essential for embryonic development."Biochem.Biophysic.Res.Comm.. 266. 569-574 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hisahara, S.et al.: "Targeted expression of baculovirus p35 caspase inhibitor in oligodendrocytes protects mice against autoimmune-mediated demyelinaion."EMBO J.. 19. 341-348 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Mcllroy, D.et al.: "An auxiliary mode of apoptotic DNA fragmentation provided by phagocytes."Genes Dev.. 14. 549-558 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hisahara,S. et al.: "Targeted expression of baculovirus p35 caspase inhibitor in oligodendrocytes protects mice against autoimmune-mediated demyelinaion."EMBO J.. 19. 341-348 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] McIlroy,D. et al.: "An auxiliary mode of apoptotic DNA fragmentation provided by phagocytes. :"Genes Dev.. 14. 549-558 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Abe,K. et al.: "Molecular and embryological characterization of a new transgene-induced null allele of mouse Brachyury locus."Mammalian Genome. 11. 238-240 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Okamoto,T.et al.: "The mouse interferon-g transgene chronic hepatitis model."Int.J.Mol.Med.:. 3. 517-520 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Yamamura,K.: "Overview of trangenic and gene knockout mice.:"Prog.Exp.Tumor Res.. 35. 13-24 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Akbar,S.et al.: "Molecular cloning of the human gene STK10 encoding LOK and comparative chromosomal mapping of the human,mouse and rat homologues."Immunogenetics. 49. 369-375 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kanai-Azuma,M.et al.: "Unique features of dendritic cells in IFN-g transgenic mice:Relevance to cancer development and therapeutic implications."Biochem.Biophis.Res.Comm.. 259. 294-299 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kondo,T.et al.: "Genomic organization and expression analysis of the mouse qkI locus."Mammal.Genome.. 10. 662-669 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Yamauchi,Y.et al.: "A novel transgenic technique that allows specific marking of the neural crest cell Iineage in mice."Dev.Biol.. 212. 191-203 (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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