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Functional characterization of homologues of repair enzyme genes for ultraviolet-induced DNA damages in higher plants.

Research Project

Project/Area Number 06660009
Research Category

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

Allocation TypeSingle-year Grants
Research Field Breeding science
Research InstitutionKagawa University

Principal Investigator

IKEDA Shigeru  Kagawa Univ., Fac., Agric., Research associate, 農学部, 助手 (90151290)

Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1995: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1994: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsDNA Repair / Rice / Gene Tranfer
Research Abstract

Two sensitive detection methods, ELISA and immuno-PCR method, utilizing monoclonal antibodies were developed to estimate the ability of rice cells to repair thymine dimers induced by ultraviolet irradiation. After rice tissues were irradiated by ultraviolet, expression of PRHs (homologues of E.coli phr ; gene for photoreactivation enzyme) and ERHs (homologues of E.coli uvrB ; gene for excision repair enzyme) were estimated by Northern blotting, and repairability of thymine dimers was estimated by the immuno-PCR method. PRH1 and ERH1 showing UV irradiation-specific expression were selected to transformate rice cells. The rice protoplasts were co-transformed with plasmids harvoring E.coli phr or uvrB,PRH1, or ERH1 and plasmids harvoring kanamycin resistance gene by electroporation. Kanamycin resistant calli showed expression of introduced repair gene or its homologue and higher repairability of thymin dimers than non-transformants. The rice protoplasts were also electroporated with yeast phr gene by the method described above, but kanamycin resistant calli showed expression of phr and the same repairablity as of non-transformants. Delesion mutant genes, phrDELTA and PRH1DELTA, lost highly conservative amino acids sequence (PIVDAAMRQL) were construced and introduced into rice protoplasts by co-transformation method described above. Kanamycin resistant calli showed expression of mutant gene and the same repairability as of non-transformants. Therefore the conclusion that PRH1 is a rice gene for photoreactivation enzyme was drawn. Unfortunately any plantlets were not regenerated from kanamycin resistant calli.

Report

(3 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (3 results)

All Other

All Publications (3 results)

  • [Publications] 池田滋: "モノクローナル抗体を用いたUV誘発DNA損傷の高感度検出" 育種学雑誌 別冊2号. 45. 213 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Ikeda Shigeru: "Highly sensitive detection of UV-induced DNA damages by using of moclonal antibodies" Breeding Journal. 45 (Suppl.2). 213 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 池田 滋: "モノクローナル抗体を用いたUV誘発DNA損傷の高感度検出" 育種学雑誌 別冊2号. 45. 213- (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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