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1993 Fiscal Year Final Research Report Summary

Production of Useful Compounds by Genetic Transformation

Research Project

Project/Area Number 03044050
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionFaculty of Pharmaceutical Sciences, University of Tokyo

Principal Investigator

SANKAWA Ushio  Professor, University of Tokyo, 薬学部, 教授 (60012613)

Co-Investigator(Kenkyū-buntansha) HAKAMATSUKA Takashi  Associate Professor, University of Tokyo, 薬学部, 教務職員 (60221488)
FUJIWARA Toru  Associate Professor, University of Tokyo, 農学部, 助手 (80242163)
SHIBUYA Masaaki  Associate Professor, University of Tokyo, 薬学部, 助手 (50170923)
FUJII Isao  Associate Professor, University of Tokyo, 薬学部, 助手 (70181302)
NAITO Satoshi  Associate Professor, Hokkaido University, 農学部, 助教授 (20164105)
KOMEDA Yoshifumi  Associate Professor, University of Tokyo, 遺伝子実験施設, 助教授 (10124215)
BEACHY Roger N.  Professor, The Scripps Institute, USA, 教授
Project Period (FY) 1991 – 1993
KeywordsChalcone Synthase / Chalcone Reductase / Oxidosqualene / Cyclase / beta-Amyrin / Cycloartenol / beta-Conglycinin / Genetic Transformation
Research Abstract

(1)The Genes of Flavonoids biosynthesis
The genes of flavonoids biosynthesis are induced by environmental stimuli. This induction was regulated at transcriptional level. In this project we focussed on the genes of chalcone synthase(CHS)and reductase(CHR)co-acting with chalcone synthase.
The cDNA of CHS was cloned from Pueraria lobata cDNA library. And the cDNA of reductase was cloned by PCR. The genome genes of CHS and CHR were cloned from genome DNA library using the each cDNA as probe.
(2)The Genes of Terpenoids Biosynthesis
The biosynthesis pathway of triterpene and sterol is dissected at cyclization of oxidosqualene in plant. Cycloartenol cyclase produces cycloartenol from oxidosqualene and leads to sterols. On the other hand, beta-amyrin cyclase, one of triterpene cyclases, cyclase oxidosqualene to beta-amyrin and leads to triterpines. In this project we focussed on the genes of cycloartenol cyclase and beta-amyrin cyclase.
These two enzymes were purified from pea, and several amino acid sequences were determined. On the basis of these sequences, the oligo nucleotides were synthesized and used as primer of PCR. The partial length cDNA was obtained, and cDNA library was screened using the obtained partial length cDNA as probe. The full length cDNA of beta-amyrin cyclase was obtained, but cDNA of cycloartenol cyclase has not been obtained full length cDNA yet.
(3)Introduction of Foreign Genes and Analysis of Promoters
The promoter region of beta-conglycinin gene which is coding the storage protein of soybean seeds is analyzed by construction of deletion clones. The sequence of "CATGCAT" is indispensable for specific expression in seeds.
In the research of introduction of foreign genes into plant, some new results about effects of adjacent sequences, effects of transferred location and effects of double transformation, were obtained.

  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] O.Nakajima: "Isolation,Sequence and Bacterial Expression of cDNA for Chalcone Syntase from the Cultured Cells of Pueraria lobata" Chem.Pharm.Bull.39. 1911-19 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Fujiwara: "Seed Specific Repression of GUS Activity in Tobacco Plants by Antisense RNA" Plant Mol.Biol.20. 1059-1069 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Fujiwara: "Expression of Soybean Seed Storage Protein Genes in Transgenic Their Effects on Expression of a Neighboring Gene and Position Dependency" Plant Cell Physiol. 34. 13-20 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Fujiwara: "Reversible Inactivation of the Nopaline Synthase Gene by Double Transformation" Plant Cell Reports. 13. 133-138 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Fujiwara: "Identification of the CATGCAT Sequence as a Functional cis Acting Element for Seed Specific Expression of a β-Conglycinin Gene" Plant Mol.Biol.(in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] O.Nakajima: "cDNA Cloning of Reductase Which is Co-acting with Chalcone Synthase in the Formation of Deoxychalcone from Cell Suspension Culture of Pueraria lobata" Biol.Pharm.Bull.(in preparation).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] O.Nakajima: "DNA Cloning of Chalcone Synthase from Pueraria lobata" Biol.Pharm.Bull. (in preparation).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] O.Nakajima: "DNA Cloning of Reductase Which is Acting in Deoxychalcone formation from Pueraria lobata" Biol.Pharm.Bull.(in preparation).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.Lee: "Promoter Activity of Pueraria lobata Chalcone Synthase Gene in Hairy Roots of Tobacco" Biol.Pharm.Bull. (in preparation).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Lee: "cDNA Cloning of b-Amyrin Cyclase from Pea" Biol.Pharm.Bull. (in preparation).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Lee: "cDNA Cloning of Cycloartenol Cyclase from Pea" Biol.Pharm.Bull. (in preparation).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] O.Nakajima, T.Akiyama, T.Hakamatsuka, M.Shibuya, H.Noguchi, Y.Ebizuka and U.Sankawa: "Isolation, Sequence and Bacterial Expression of cDNA for Chalcone Syntase from the Cultured Cells of Pueraria lobata" Chem.Pharm.Bull.39. 1911-1993 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Fujiwara, P.A.Lessard, R.N.Beachy: "Seed Specific Repression of GUS Activity in Tobacco Plants by Antisense RNA" Plant Mol. Biol.20. 1059-1069 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Fujiwara, R.N.Beachy: "Expression of Soybean Seed Storage Protein Genes in Transgenic Plants ; Their Effects on Expression of a Neighboring Gene and Position Dependency" Plant Cell Physiol. 34. 13-20 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Fujiwara, P.A.Lessard and R.N.Beachy: "Reversible Inactivation of the Nopaline Synthase Gene by Double Transformation" Plant Cell Reports. 13. 133-138 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Fujiwara and R.N.Beachy: "Identification of the CATGCAT Sequence as a Functional cis Acting Element for Seed Specific Expression of a beta-Conglycinin Gene" Plant Mol. Biol.(in press). (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] O.Nakajima, M.Shibuya, Y.Ebizuka and Ushio Sankawa: "cDNA Cloning of Reductase Which is Co-acting with Chalcone Synthase in the Formation of Deoxychalcone from Cell Suspension Culture of Pueraria lobata" Biol.Pharm.Bull.(in preparation).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] O.Nakajima, M.Shibuya, Y.Ebizuka and Ushio Sankawa: "DNA Cloning of Chalcone Synthase from Pueraria lobata" Biol.Pharm.Bull. (in preparation).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] O.Nakajima, M.Shibuya, Y.Ebizuka and Ushio Sankawa: "DNA Cloning of Reductase Which is Acting in Deoxychalcone formation from Pueraria lobata" Biol.Pharm.Bull. (in preparation).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J.Lee, O.Nakajima, M.Shibuya, Y.Ebizuka and Ushio Sankawa: "Promoter Activity of Pueraria lobata Chalcone Synthase Gene in Hairy Roots of Tobacco" Biol.Pharm.Bull. (in preparation).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Lee, I.Abe, M.Kusano, M.Shibuya, Y.Ebizuka and Ushio Sankawa: "cDNA Cloning of beta-Amyrin Cyclase from Pea" Biol.Pharm.Bull. (in preparation).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Lee, I.Abe, M.Kusano, M.Shibuya, Y.Ebizuka and Ushio Sankawa: "cDNA Cloning of Cycloartenol Cyclase from Pea" Biol.Pharm.Bull. (in preparation).

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

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Published: 1995-02-07  

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