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1999 Fiscal Year Final Research Report Summary

Mapping and cloning of the Da mutant gene defective dorsal-ventral patterning

Research Project

Project/Area Number 10680686
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Developmental biology
Research InstitutionNAGOYA UNIVERSITY

Principal Investigator

OZATO Kenjiro  Nagoya University, Bioscience Center, Profesor, 生物分子応答研究センター, 教授 (90026790)

Co-Investigator(Kenkyū-buntansha) KIMURA Minoru  Tokai University, Medical School, Professor, 医学部, 教授 (10146706)
Project Period (FY) 1998 – 1999
Keywordsmedaka / morphogenesis / dorsal-ventral patterning / positional cloning
Research Abstract

Formation of dorsal-ventral patterning is one of the major issues of vertebrate development. Medaka Da mutant expresses a phenotype in which structures characteristic to the dorsal region are ventralized. In the present study, we attempted to identify the Da gene by positional cloning.
1) Construction of the linkage map and mapping of the Da gene.
The genetic markers we used allowed us to identify 26 linkage groups in the medaka genome, in which the Da gene was mapped on the linkage group VIII. The physical distance from Da to two markers between which put Da was 144kb and 360kb, respectively. These distances were close enough to conduct chromosomal walking.
2) Construction of Cosmid library
We constructed a Cosmid library using the medaka HNI-I strain, which consisted of 120,000 clones (corresponding to 6 haploid genomes) with an average insert-length of 40.2kb. We successfully obtained Cosmid clones containing two markers close to Da by screening the library.
3) Identification of Da in one BAC clone.
Using Cosmid and BAC clones, we determined a minimum region containing Da, demonstrating that Da was contained in a range of 70kb to 250kb. Thus, it was expected that the Da gene was covered by one BAC clone.

  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] Hamada, K.: "Usefulness of the medaka B-actin promoter investigated using a mutant GFP reporter gene in transgenic medaka (Oryzias latipes)"Mol. Marine Biol. Biotech. 7・3. 173-180 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Niwa, K.: "Transplantation of blastula nuclei, to non-enucleated eggs in the medaka, Oryzias latipes"Develop. Growth Differ.. 41・2. 163-172 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sasado, T.: "Expression of murine early embryonic antigens, SSEA-1 and antigenic determinant of EMA-1, inevbryos and ovarian follicles of a teleost medaka (Oryzias latipes)"Develop. Growth Differ.. 41・3. 293-302 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ohtsuka, M.: "Construction of Linkage Map of the Medaka(Oryzias latipes) and Mapping of the Da Mutant Locus Defective in Dorsoventral Patterning"Genome Res.. 9. 1277-1287 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hamada, K. et al.: "Usefullness of medaka β-actin promotor investigated using a mutant GFP reporter gene in transgenic medaka (Oryzias latipes)"Mol Marine Biol Biotech. 7 (3). 173-180 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Niwa, K. et al.: "Transplantation of blastula nuclei to non-enucleated eggs in medaka, Oryzias latipes"Develop Growth Differ. 41 (2). 163-172 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sasado, T. et al.: "Expression of murine early embryonic antigens, SSEA-1 and antigenic determinant of EMA-1, in embryos and ovarian follicles of a teleost, medaka (Oryzias latipes)"Develop Growth Differ. 41 (3). 293-302 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ohtsuka, M. et al.: "Construction of a linkage map of the medaka (Oryzias latipes) and mapping of the Da mutant locus defective in dorsoventral patterning"Genome Res.. 9. 1277-1287 (1999)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2001-10-23  

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