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THE STUDY OF HOMOLOGOUS RECOMBINATION AND GENE REPAIR IN MOUSE SOMATIC CELLS.

Research Project

Project/Area Number 07640823
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 遺伝
Research InstitutionTOKAI UNIVERSITY

Principal Investigator

GONDO Yoichi  TOKAI UNIVERSITY,THE INSTITUTE OF MEDICAL SCIENCES,ASSOCIATE PROFESSOR, 総合医学研究所, 助教授 (40225678)

Co-Investigator(Kenkyū-buntansha) SAITOH Yasushi  THE INSTITUTE OF MEDICAL SCIENCES TOKAI UNIVERSITY RESEARCH ASSOCIATE, 総合医学研究所, 助手 (70287100)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1996: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1995: ¥1,300,000 (Direct Cost: ¥1,300,000)
Keywordshomologous recombination / somatic cells / gene conversion / mouse / tissue culture cells / DNA repair / molecular genetics / 高等動物 / 分子遺伝子 / ジーンターゲッテイング / マウスゲノム / 不等価組換え
Research Abstract

The major objective of this study is to establish a sensitive method of detecting chromosomal recombinations and to analyze recombinational mechanisms at the molecular level. Mouse embryonic stem (ES) cell lines that carried a neo-tk cassette at the p53 locus on the chromosome 11 were established by using gene targeting (Gondo et al., Biochem.Biophys.Res.Commun., 202 : 830-837,1994). These ES cell lines were, thus, heterozygotes carrying the wild-type (W) and neo-tk (T) alleles at the p53 locus. When W/T heterozygous cells were treated with GANC,recombinational repair genotype, namely, W/W homozygotes were detected. Reciprocally, T/T homozygotes were also isolated with a high concentration of G418. In order to investigate the molecular genetic changes at and around the recombinational repair region, we also established W/T heterozygotes by using TT2 ES cells which derived from a CBAXC57BL/6 F1 mouse. Preliminary studies indicated approximately a half of chromosome 11 markers were heter … More ozygous in TT2. By using these polymorphic markers of chromosome 11 and identified recombinants of W/T to W/W or T/T,it is now possible to analyze whether the recombinational repair was manifested by 1) gene conversion with a narrow sense, 2) mitotic crossing-over between two homologous, or 3) uniparental disomy-type irregular chromosomal segregation.
We also developed a new marker gene, Eco-gpt, in addition to HSV-tk to enhance the sensitivity as well as to reduce the labor-intensive tasks of the selection system. We have established ES lines carrying neo-gpt cassette (G) allele and confirmed the genotypes were W/G heterozygotes (Ito, Gondo et al., in preparation). W/W homozygotes were also isolated from W/G ES cells by 6-tg selection. Having two distinctive negatively selectable marker genes like HSV-tk and Eco-gpt, it is plausible to construct T/G heterozygotes. For instance, neo-tk integration to one allele and puromycin^R-gpt integration to the other should provide such heterozygotes. It will provide a new tool of the selection for recombinational repair between two heterozygous homologues to both directions ; from G/T to T/T and to G/G.The tagging of HSV-tk and Eco-gpt should be applicable to any part of mammalian genome including human when a tissue culture system is available. Thus by using the established system homologous recombination is now feasible to be investigated at the molecular level to elucidate the enzymatic process at any genes and species. Less

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] T.Kitamoto,Y.Gondo,et al.: "Humanized prion protein knock-in by Creinduced site-specific recombination in the mouse." Biochem.Biophys.Res.Commun.222. 742-747 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Shibata,Y.Gondo,et al.: "Inactive allele-specific methylation and chromatin structure of the imprinted gene U2af1-rs1 on mouse chromosome 11." Genomics. 35. 248-252 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Norimura,Y.Gondo,et al.: "p53-dependent apoptosis suppresses radiation-induced teratogenesis." Nature Medicine. 2. 577-580 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Y.Gondo,Y.Shioyama,et al.: "A novel positive detection system of in vivo mutations in rpsL(strA)transgenic mice." Mutation Res.360. 1-14 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 権藤洋一、勝木元也: "遺伝毒性検定のためのトランスジェニックマウス." 組織培養. 22(13). 522-525 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 権藤洋一: "発がんリスク検定のためのトランスジェニックマウス." Pharma Sandoz. 3(21). 22-23 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 権藤洋一、勝木元也: "11.4 DNA変異を検出する方法." 講談社サイエンティフィク, 11 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tetsukuki Kitamoto, Kenji Nakamura, Kazuki Nakao, Satoshi Shibuya, Ryong-Woon Shin, Yoichi Gondo, Motoya Katsuki and Jun Tateishi: "Humanized prion protein knock-in by Cre-induced site-specific recombination in the mouse." Biochem.Biophys.Res.Commun. 222. 742-747 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideo Shibata, Kiyoshi Yoshino, Shoichi Sunahara, Yoichi Gondo, Motoya Katsuki, Takayuki Ueda, Mamoru Kamiya, Masami Muramatsu, Yasufumi Murakami, Iveta Kalcheva, Christoph Plass, Verne M.Chapman and Yoshihida Hayashizaki: "Inactive allele-specific methylation and chromatin structure of the imprinted gene U2af1-rs1 on mouse chromosome 11." Genomics. 35. 248-252 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yoichi Gondo, Yoshiyuki Shioyama, Kazuki Nakao, Motoya Katsuki: "A novel positive detection system of in vivo mutations in rpsL (strA) transgenic mice." Mutation Res.360. 1-14 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Toshiyuki Norimura, Satoshi Nomoto, Motoya Katsuki, Yoichi Gondo and Sohei Kondo: "p53-dependent apoptosis suppresses radiation-induced teratogenesis." Nature Medicine. 2. 577-580 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yoichi Gondo and Motoya Katsuki: "Transgenic system for the genotoxicity test. (in Japanese)" The Tissue Culture. 22 (13). 522-525 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yoichi Gondo: "Transgenic mouse for the carcinogenicity test. (in Japanese)" Pharma Sandoz. No.3 (21). 22-23 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yoichi Gondo and Motoya Katsuki: Detection methods of DNA mutations (in Japanese) in Antimutagen, Anticarcinogenand their screening methods (ed.by Y.Kuroda). Kodansha Scientific Pless, Tokyo, Japan, 387-397 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Kitamoto,Y.Gondo,et al.: "Humanized prion protein knock-in by Creinduced site-specific recombination in the mouse." Biochem.Biophys.Res.Commun.222. 742-747 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] H.Shibata,Y.Gondo,et al.: "Inactive allele-specific methylation and chromatin structure of the imprinted gene U2af1-rs1 on mouse chromosome 11." Genomics. 35. 248-252 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Norimura,Y.Gondo,et al.: "P53-dependent apoptosis suppresses radiation-induced teratogenesis." Nature Medicine. 2. 577-580 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Gondo,Y.Shioyama,et al.: "A novel positive detection system of in vivo mutations in rPSL (strA) transgenic mice." Mutation Res.360. 1-14 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 権藤洋一、勝木元也: "遺伝毒性検定のためのトランスジェニックマウス." 組織培養. 22(13). 522-525 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 権藤洋一: "発がんリスク検定のためのトランスジェニックマウス." Pharma Sandoz. 3(21). 22-23 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 権藤洋一、勝木元也: "11.4DNA変異を検出する方法." 講談社サイエンティフィク, 11 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y. Gondo, et al.: "A novel positive detection system of in vivo mutations in rpsL (StrA) transgenic mice." Mutation Res.(in press). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] 権藤洋一 他: "「抗変異原・抗発ガン物質とその検索法」11.4 DNA変を検出する方法" 講談社サイエンティフック, 11 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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