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1990 Fiscal Year Final Research Report Summary

Gene Structure and Fruits Specific Expression of Melon Serine Proteinase

Research Project

Project/Area Number 01560096
Research Category

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

Allocation TypeSingle-year Grants
Research Field 応用生物化学・栄養化学
Research InstitutionKobe University

Principal Investigator

YAMAGATA Hiroshi  Kobe Univ. Faculty of Agriculture, Associate Professor, 農学部, 助教授 (00159203)

Project Period (FY) 1989 – 1990
KeywordsMelon / Serine proteinase / Fruit enzyme / Protein synthesis / cDNA cloning
Research Abstract

Melon serine proteinase (MSP) is a major protein in the fruits of melon. The time and the site of MSP synthesis is restricted to the developing young fruits. The goal of this project is to clear the molecular mechanisms of the gene expression and the protein synthesis of the MSP.
The tissue blotting assay and the incorporation of the labeled amino acid into the protein in the tissue block suggested that the MSP is synthesized in the seed sac or its peripheral tissue.
Using guanidinium isothiocyanate and ultra centrifugation, a mRNA which had a high template activity of protein synthesis in vitro can be isolated. The differences were observed among the in vitro products of mRNA from the several parts in the fruits. A cDNA was synthesized from the mRNA isolated from the center parts of the fruits and ligated to the lambda gt 11 arm and then in vitro packaged phage was introduced into E. coli Y1090R-. This constructed cDNA library was screened using anti-MSP rabbit antibody and 6 positive clones could be isolated. The length of the 5 cDNAs of these clones was 700 to 1,900 bp. The longest clone, lambda MSP 28 had a cDNA whose length is thought to be near full-length of the MSP mRNA. These 5 cDNA clone were subcloned into phagemid pBluescript and their sequencing are now being carryied out.
On the other hand, genome DNA was extracted from young melon leaves using CTAB, and was partially hydrolyzed by Sau 3A. Resulting the DNA of 10 to 20 kb was ligated to the EMBL 3 arm and the genome libraly was prepared. In the following research, the structure of the isolated cDNA and the MSP gene will be analyzed and clear the cis elements responsible for the fruit specific expression of the MSP gene.

  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] 山形 裕士,上野 修治,岩崎 照雄: "Isolation and Characterization of a Possible Native Cucumisin from Developing Melon Fruits and Its Limited Autolysis to Cucumisin." Agric.Biol.Chem.53. 1009-1017 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 花森 清茂,山形 裕士,岩崎 照雄: "メロンセリンプロテア-ゼの果実内動態" 日本農芸化学会誌. 64. 285 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山形 裕士,花森 清茂,岩崎 照雄: "Biosynthesis and Localization of Serine Proteinase in the Developing Melon Fruits." Plant Physiol.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山形 裕士,大西 達司,増沢 拓也,岩崎 照雄: "cDNA cloning of an mRNA Encoding a Serine Proteinase in Melon Fruits." Plant Mol.Biol.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamagata Hiroshi: "Isolation and characterization of a possible native cucumisin from developing melon fruits and its limited autolysis to cucumisin." Agric. Biol. Chem.53. 1009-1017 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hanamori Kiyoshige: "Change of serine proteinase in the melon fruits." Nippon Nogeikagaku Kaishi. 64. 285 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamagata Hiroshi: "Biosynthesys and localization of serine proteinase in the developing melon fruits."

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamagata Hiroshi: "cDNA cloning of and mRNA encoding a serine protenase of melon fruits."

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

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Published: 1993-08-12  

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