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Analysis of the genes for porphyrin biosynthetic pathway in higher plants utilizing the light-sensitive mutants of Escherichia coli

Research Project

Project/Area Number 11480201
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field Molecular biology
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

INOKUCHI Hachiro  Dept.of Biophys., Gard.Sch.Sci.Kyoto Univ. (Professor), 大学院・理学研究科, 教授 (20028195)

Co-Investigator(Kenkyū-buntansha) NAKAHIGASHI Kenji  Dept.of Biophys., Gard.Sch.Sci.Kyoto Univ.(Assistant Professor), 大学院・理学研究科, 助手 (70322740)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 2000: ¥4,600,000 (Direct Cost: ¥4,600,000)
Keywordsrice (Oryza sativa) / soybean (Glycine max) / hemF gene of E.coli / cucumber (Cucumis sativus) / thymidylate synthase / spinach (Spinacia oleracea) / ferrochelatase / protoporphyrinogen IX oxidase / ポルフィリン / PPOX / CPOX / シロイヌナズナ / キューリ / ダイズ / 大腸菌
Research Abstract

1) By taking advantage of the ability of such a cDNA to complement the thymine auxotrophy of a mutant of Escherichia coli, a cDNA clone for thymidylate synthase (TS) of rice (Oryza sativa L.indica) was isolated. [Kanjo & Inokuchi, Plant Physiol. (Gene Register), 120, 634 (1999)]
2) A cDNA clone for protoporphyrinogen IX oxidase of soybean (Glycine max was isolated by functional complementation against the poor growth of BT3 delta hemG : : Km^R cells. Nucleotide sequence analysis of the cDNA insert revealed that the insert was 1745 bp in length. It included a single open reading frame that encoded a protein consisting of 502 amino acid residues. The deduced amino acid sequence of PPOX from soybean was 67 % homologous to that of mitochondrial PPOX from tobacco. It shares some amino acid residues at the N-terminus speculated to be a mitochondrial translocation signal. [Kanjo et al., Plant Physiol. (Gene Register), 121, 1384 (1999)]
3) We examined poessible alternate pathways for the oxidati … More on of protoporphyrinogen IX to protoporphyrin IX, by isolating and investigating E.coli mutants that can still grow normally when the hemG gene is disrupted. It was suggested that the E.coli aerobic coproporphyrinogen oxidase has an intrinsic capacity to oxidize not only coproporphyrinogen III but also protoporhyrinogen IX.[Narita et al., Mol. Gen. (Genet., 261, 1012-1020 (1999)]
4) A putative mature region of a cucumber (Cucumis sativus) ferrochelatase cDNA (hemH) was overexpressed in E.coli, purified to homogenity and examined its enzymatic properties. Immunolocalization of the enzyme examined with antibody demonstrated that the ferrochelatase was present in both hypocotyls and roots but hardly in cotyledons of cucumber. [Suzuki et al., Plant Cell Physiol., 41, 192-199 (2000)]
5) We reported the first cloning of spinach (Spinacia oleracea) plastidal PPOX cDNA and analysis of the N-terminal sequence in mature plastidal PPOX purified from spinach chloroplast and discussed the mechanism of the transport of plastidal PPOX into chloroplasts. [Che et al., Plant Physiol., 124, 59-70 (2000)] Less

Report

(3 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] N.Kanjo & H.Inohuchi: "Nucleotide sequence of a CDNA Clone encoding thymidylate synthase from rice"Plant Physiol.(Gene Register). 120. 634-634 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Narita,S.Taketani & H.Inokuchi: "Oxidation of Protoporphysinogen IX mediated by the E.Coli aerobic coprosporphysinogen Oxidase"Molec.Gen.Genet.. 261. 1012-1020 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Kanjo et al.,: "Nucleotide Sequence of a cDNA clone encoding ptoto porphysinogen IX oxidase from soybean"Plant physiol.(Gene Register). 112. 1384-1384 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Suzuki et al.,: "Overexpression,Enzymatic properties and tissue Localization of a ferrochelatase of cucumber"Plant Cell physiol.. 41. 192-199 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] F.Che et al.,: "Molecular characterization subcellular localization PPOX in spinach chloroplasts"Plant Physiol.,. 124. 59-70 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Kanjo & H.Inokuchi: "Nucleotide sequence of a cDNA clone encoding thymidylate synthase from rice."Plant Physiol. (Gene Register). 120. 634 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Narita, S.Taketani & H.Inokuchi: "Oxidation of protoporphyrinogen IX mediated by the Echerichia coli aerobic coproporphyrinogen oxidase."Molec.Gen.Genet.. 261. 1012-1020 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Kanjo, S.Nishio, S.Narita, K.Oeda & H.Inokuchi: "Nucleotide sequence of a cDNA clone encoding protoporphyrinogen IX oxidase from soybean."Plant Physiol. (Gene Register). 112. 1384 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Suzuki, T.Masuda, H.Inokuchi, H.Shimada, H.Ohta & K.Takamiya: "Overexpression, Enzymeatic properties and tissue localization of a ferrochelatase of cucumber."Plant Cell Physiol.. 41. 192-199 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] F.Che, N.Watanabe, M.Iwano, H.Inokuchi, S.Takayama, S.Yoshida & A.Isogai: "Molecular characterization subcellular localization of protoporphyrinogen oxidase in spinach chloroplasts."Plant Physiol.. 124. 59-70 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Kanjo & H.Inokuchi: "Nucleotide Sequence of cDNA clone encoding thymidylate synthase from sics."Plant Physcol(Gene Regioter). 120. 634-634 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] S.Nasita et al.,: "Oxidation of Protoporphysinogen IX mediated by the E.coli aerobic coproposphysinogen oxidase"Molec.Gen.Genet.. 261. 1012-1020 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] N.Kanjo et al.,: "Nucleotide sequence of cDNA clone encoding protoporphysinogen IX oxidase from soybean"Plant Physiol.(Gene Register). 112. 1384-1384 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] T.Suzuki et.al.,: "Ouerepression, Enzymatic Propesties and tissue localization of a ferrochelatase of cucumber"Plant Cell Physiol.,. 41. 192-199 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] F.Che et.al.,: "Molecular characterization subcellular localization PPOX in spinach chlosoplasts"Plant Physiol.,. 124. 59-70 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] S.Narita: "Oxidation of protoporphyrinogen IX mediated by the Escherichia coli aerobic coproporphyrinogen oxidase"Molec. Gen. Genet.,. 261. 1012-1020 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] N.Kanjo: "Nucleotide sequence of cDNA clone encoding ferrochelatase from rice"Plant Physiol. (Gene Register). 115. 1729 (1998)

    • Related Report
      1999 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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