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DEVELOPMENTAL GENETIC ANALYSIS OF MAMMALIAN GENOME FUNCTIONS

Research Project

Project/Area Number 05671885
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Human genetics
Research InstitutionKumamoto University

Principal Investigator

ABE Kuniya  Kumamoto University, Associate professor, 医学部, 助教授 (40240915)

Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1993: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsDEVELOPMENT / YAC CLONING / TRANSGENIC MOUSE / 4T / t COMPLEX / 発生分化 / H-2 / tコンプレックス / YAC クローニング / トランスジェニックマウス / ゲノムクローニング / H-2K
Research Abstract

Positional cloning approach has been successful in isolation of mutated genes of various kinds, including genes responsible for tumorigenesis. However, despite such successes and recent progress in gene mapping and genomic cloning technologies, the identification and isolation of mutated genes still remains formidable task. In cases no gross genomic rearrangements e.g.translocation or deletion are found, it is very difficult to pinpoint a responsible gene within a large genomic segment. Yhus, there is an increasing demand for a method to functionally characterize the cloned genomic DNA.In this study, a method that allows rapid recovery of transcribed sequences from large cloned genomic DNA has been established. Also, in order to functionally characterize large genomic DNA,a technique to generate transgenic mice carrying YAC DNA is being developed.
The T/t complex of mouse is a large genetic region on proximal half of chromosome 17 that carry a number of loci affecting embryogenesis and … More germ cell functions. We have been analyzing this region, attempting to ultimately isolate genes responsible for these mutations. Among these, we have special interests in mutations such ast^<omega5>, a recessive postimplantation lethal, showing defects in embryonic ectoderm formation ; qk (quaking), a neurological mutant. We have mapped t^<omega5> within mouse MHC region, H-2 complex. t^<omega5> was found to be very close to the H-2K gene with a possible genetic distance less than 0.1cM.We have cloned genomic region spanning -200kb around the K gene into cosmid contigs and have searched genes within the cloned region. Now, we extended the cloned area by isolating several YAC clones that cover -800kb from the K-gene toward CryA1 marker. Genetic analysis suggests that the mutation lies within the cloned DNA,while no structural alterations are found so far in this region. Twenty two novel genes have been found in this cloned genomic area by using our' screening in solution' method. Also, to find a responsible gene, a technique to introduce YAC DNA into germ line has been applied. We first established a protocol for isolation and purification of large DNA fragment without breakage, and thus isolated 650 kb YAC from t^<omega5> region was injected into mouse fertilized eggs. After transferring 320 injected eggs to pseudopregnant foster, 39 pups were born. Of which 3 mice carry at least a part of the YAC vector sequence, suggesting transgene integration. Genomic DNA from these putative transgenic mice are now being characterized in detail. Less

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (20 results)

All Other

All Publications (20 results)

  • [Publications] Nagata,T: "Physical mapping of the retinoid X receptor β in mouse and man." Immunogenetics. (in press.).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mantani,A.: "A novel isoform of neurofibromatosis type-1m RNA and a switch of isoforms during murine cell differentiation and proliferation." Gene. 148. 245-251 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kimura,S.: "Improvement of germ line transmission by targeting β-galactosidase to nuclei in transgenic mice." Develop.Growth & Diff.36. 521-528 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ando,A.: "Cloning of a new kinesin-related gene located at the centromeric end of the human MHC region." Immunogenetics. 39. 194-200 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Wada,M.: "Chimeric YACs were generated at unreduced rates in conditions that suppress coligation." Nucleic Acids Research. 22. 1651-1654 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Toyonaga,T.: "Chronic active hepatitis in transgenic mice expressing interferon-γ in the liver." Proc.Natl.Acad.Sci.USA.91. 614-618 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 阿部訓也: "マニュアル疾患モデルマウス.Wnt遺伝子関連のノックアウトマウス" 中山書店(東京), 70-74 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Nagata, T., Weiss, E.H., Abe, K., Kitagawa, K., Ando, A., Yara-kikuti, Y., Seldin.M.F., Ozato, K., Inoko, H., and Takeo, M.: "Physical mapping of the retinoid X receptor beta in mouse and man." Immunogenetics. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mantani, A., Wakasugi, S., Yokata, Y., Abe, K., Ushio, Y.and Tamamura, K.: "A novel isoform of neurofibromatosis type-1 mRMA and a switch of isoforms during murine cell differentiation and proliferation." Gene. 148. 245-251 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kimura, S., Niwa, H., Moriyama, M., Araki, K., Abe, K., Miike, T.and Yamamura, K.: "Improvement of germ line transmission by targeting beta-galactosidase to nuclei in transgenic mice." Develop.Growth epsilon Diff. 36. 521-528 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ando, A., Kikuti, Y.Y., Kawata, H., Okamoto, N., Imai, T., Eki, T., Yokoyama, K., Soeda, E., Ikwmura, T., Abe, K.and Inoko, H.: "Cloning of a new kinesin-related gene located at the centromeric end of the human MHC region." Immunogenetics. 39. 194-200 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Wada, M., Abe, K., Okumura, K., Taguchi, H., Kohno, K., Imamoto, F., Schlessinger, D.and Kuwamo, M.: "Chimeric YACs were generated at unreduced rates in conditions that suppress coligation." Nucleic Acids Research. 22. 1651-1654 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Toyonaga, T., Hino, O., Sugai, S., Wakasugi, S., Abe, K., Shichiri, M., and Yamamura, K.: "Chronic active hepatitis in transgenic mice expressing interferon-gamma in the liver." Proc.Natl.Acad.Sci.USA. 91. 614-618 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Nagata,T: "Physical mapping of the retinoid X receptor β in mouse and man." Immunogenetics in press.(1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Mantani,A.: "A novel isoform of neurofibromatosis type-1 mRNA and a switch of isoforms during murine cell differentiation and proliferation." Gene. 148. 245-251 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Kimura,S.: "Improvement of germ line transmission by targeting β-galactosidase to nuclel in transgenic mice." Develop.Growth & Diff.36. 521-528 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ando,A.: "Cloning of a new kinesin-related gene located at the centromeric end of the human MHC region." Immunogenetics. 39. 194-200 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Wada,M.: "Chimeric YACs were generated at unreduced rates in conditions that suppress coligation." Nucleic Acids Research. 22. 1651-1654 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Toyonaga,T.: "Chronic active hepatitis in transgenic mice expressing interferon-γ in the liver." Proc.Natl.Acad.Sci.USA.91. 614-618 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ando,A.: "Cloning of a new Kinesin-related gene located at The centromeric and of The human MHC rigion" Immunogenetics. 39. 194-200 (1994)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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