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

Transgenic animal model showing genomic instabilities like human hypervariable minisatellite and its applications in forensic genetics.

Research Project

Project/Area Number 18390205
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Legal medicine
Research InstitutionKyoto University

Principal Investigator

TAMAKI Keiji  Kyoto University, Graduate School of Medicine, Dept. Forensic Med. and Molecualor Pathology, Professor (90217175)

Co-Investigator(Kenkyū-buntansha) TODO Takeshi  Osaka University, Graduate School of Medicine, Faculty of Medicine, Professor (90163948)
TAKEDA Shunichi  Kyoto University, Faculty of Medicine, Professor (60188191)
YAMAMOTO Toshimichi  Nagoya University, Graduate School of Medicine, Faculty of Medicine, Associate Professor (50260592)
TSURUYAMA Tatsuaki  Kyoto University, Faculty of Medicine, Associate Professor (00303842)
OKUNO Tomoko  Kyoto University, Faculty of Medicine, Assistant Professor (30288386)
Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥16,500,000 (Direct Cost: ¥14,700,000、Indirect Cost: ¥1,800,000)
Fiscal Year 2007: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
Fiscal Year 2006: ¥8,700,000 (Direct Cost: ¥8,700,000)
Keywordsminisatellite / Medaka genome / animal model
Research Abstract

Since Sir Alec Jeffreys' initial discovery of tandemly repeated DNA units varying amongst different individuals in 1984, a growing body of evidence has come to describe the element of human hypervariable minisatellites. Decades of active research dedicated solely to these tandem repetitive units have produced a vast amount of insight into their underlying molecular basis; however, the genomic dynamics of minisatellites at meiosis in germ cells has long remained an elusive issue. In the present study, Chyzias latipes (or Medaka) were selected to examine genomic instability in minisatellites owing to the inherent difficulty of human testing.
Information retrieved from the Medaka Genome Database identified 30,504 VNTR (variable number of tandem repeats) sequences within the8chromosomes comprising the medaka genome. These results are loosely comparable to that of the human genome. Conversely, the number of expected hypervariable minisatellites-using HistoryR and GC content as an index謡as su … More bstantially lower in medaka. To initiate the study, 174 minisatellite loci were selected on 4 medaka chromosomes. Of these, 31 loci were interrogated for polymorphisms via polymerase chain reaction (PCR) using DNA derived from Oryzias latipes and Cab as templates. Preliminary screening efforts revealed multiple alleles at four of these loci, for which sequences were subsequently determined.
Next, trans-minisatellite medaka were established by introducing the short allele of human minisatellite B6.7 into fertilized medaka eggs. 22 individual fish survived to adulthood (GO), of which the presence of B6.7 was only confirmed in a small fraction (n=4). In the following generation (F1), hundreds of fish were screened, but the search ended in zero positive confirmations of B6.7 alleles. Consequently, the scope of the screening was reconstituted to include the flanking sequences of the human alleles, and B6.7 was reinserted into a new batch of fertilized medaka eggs. This produced 25 fish (GO), in which one was positive for the B6.7 allele. This fish was crossed with non-injected GO fish to establish the F1 generation. In total, 325 F1 fish were born and B6.7 was retained in the genome of 4 of these fish. These 4 fish were back-crossed to produce the F2 generation. The retention rate of B6.7 in F2 fish drastically increased to 45%. Following this, F2 were further back-crossed to establish the F3 generation. At present, F3 fish are in the process of being self-mated (F3 with F3) in order to establish B6.7 homozygotes. While our preliminary results showed 112/257 eggs positive for B6.7 in heterozygous (F2) fish, no length variance, nor mutations, were noted in these alleles. Less

Report

(3 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • Research Products

    (7 results)

All 2007

All Journal Article (4 results) (of which Peer Reviewed: 1 results) Presentation (2 results) Book (1 results)

  • [Journal Article] メダカゲノム内のミニサテライトに関する検討2007

    • Author(s)
      奥野知子
    • Journal Title

      DNA多型 15

      Pages: 27-30

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Minisatellite in the Medaka genome2007

    • Author(s)
      Okuno T., et al.
    • Journal Title

      DNA polymorphism 15

      Pages: 27-30

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Minisatellite and microdstellite DNAtyping analysis.2007

    • Author(s)
      Tamaki K.
    • Journal Title

      Molecular Forensics. John Wiley&Sons Ltd.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] メダカゲノム内のミニサテライトに関する検討2007

    • Author(s)
      奥野知子
    • Journal Title

      DNA多型 (印刷中)

    • Related Report
      2006 Annual Research Report
  • [Presentation] メダカゲノム内のミニサテライトに関する検討2007

    • Author(s)
      奥野知子
    • Organizer
      日本DNA多型学会第16回学術集会
    • Place of Presentation
      広島県福山市
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Minisatellite in the Medaka genome2007

    • Author(s)
      Okuno T., et al.
    • Organizer
      DNA polymorphism
    • Place of Presentation
      Fukuyama
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Book] Minisatellite and microsatellite DNA typing analysis. Molecular Forensics2007

    • Author(s)
      Tamaki K
    • Publisher
      John Wiley & Sons Ltd.
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary

URL: 

Published: 2006-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi