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Gene Cloning and Expression of Recombination Gene of Buckwheat Protein FE25

Research Project

Project/Area Number 06680024
Research Category

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

Allocation TypeSingle-year Grants
Research Field 家政学
Research InstitutionEhime University

Principal Investigator

TAKUMI Kenji  Ehime University, Faculty of Education, Professor, 教育学部, 教授 (90035428)

Co-Investigator(Kenkyū-buntansha) KOGA Tetsuro  Tokushima University, School of Medicine, Department of Nutrition, Tutor, 医学部, 助手 (20093859)
Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1994: ¥1,200,000 (Direct Cost: ¥1,200,000)
Keywordsbuchwheat storage protein / FE25 Globulin / Eco RI and Sal-1 restriction enzymes / pCU8 plasmid / Sucrose density gradient / poly (A) -RNA / cDNA encoding FE25 globulin / cGNA library / 25kDaサブユニット / Sau3AI制限酵素 / λEMBL3 / 15〜20Kbp DNA新片
Research Abstract

This study has been attempted for the purpose of development an improved geno-type of buckwheat grain, by which it could be expected that some intrinsic faults of the grain, including rough-textured nature, or allergenic property would be removed or weaken. Thus to construct and expression of cDNA encoding buckwheat globulin (25FE), which is the major storage protein of the grain, is a main goal of the study.
In 1994, total seed protein RNA was prepared from the endosperm of 10-day after flower and then poly (A) -enriched RNA was isolated from the RNA by two cycles of adsorption and elution from oligo (U) -agarose column. Starting with 25 mu g of poly (A) -RNA,first- and second-strand cDNA reactions were performed by using avian myeloblastosis virus (AMV) reverse transcriptase.
In 1995, the constructed cDNA was digested with both Eco R1 and Sal-1, and cDNAs larger than 500 bp were purified on a Sepharose 4B minicolumn and inserted into pCU8 for expression. The colonies were reprica plated onto nitrocellulose filters and the library was screened by anti-buckwheat globulin FE25 antibody following with ^<125>I-labeled goat anti-rabbit IgG.Only 4 colonies were detected that reacted specifically with the antisera. At present we are investigating tha colonies for the identification of cDNA encoding FE25 globulin.

Report

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

Research Products

(11 results)

All Other

All Publications (11 results)

  • [Publications] T.Koga and K.Takumi: "Isolation and charonterization of chreyporin-like proteins from vibro vilficus." Microbiol.Immunol.38. 931-936 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T.Koga and K.Takumi: "Comparisoy of chemical properties of Poriu-like proteins of V.Parahoemolyticus." Gen.Appl.Microbiol.40. 121-125 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] K.Takumi and T.Koga: "Nutrient sborvation induce cross-Protection against heat,asmotic or H_2O_2 challenge in V.Parahoemolyticus." Microbiol.Immunol.39. 22-27 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kenji Takumi,et al.: "Immunological Cross-reactivity between globulins from buckwheat and ludege seeds" 59. 1971-1972 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] K.Takumi et al.: "Polypeptide compositions and antigenic homologous among prolamins from Italian,Common,and Japanese millet Cultivars." J.Sci,Food Agric.72. 141-147 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] KOGA T.and TAKUMI K.: "Isolation and characterization of three porin-like proteins from Vibrio vulnificus." Microbiol. Immunol.38. 931-936 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] KOGA T.and TAKUMI K.: "Comoparison of chemical permeable properties of porin-like proteins a and b of V.parahaemolyticus." Gen. Appl. Microbiol.40. 11121-125 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] TAKUMI K.and KOGA T.: "Nutrient starvation induce crossprotection against heat, osmotic or H_2O_2 challenge in V.parahaemo lyticus." Microbiol. Immunal.39. 22-27 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] TAKUMU K.and KOGA T.: "Immunological cross-reactions between globulins from buckwheat and Indigo seeds." Biosci. Biotechnol. Biochem.59. 1971-1972 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] TAKUMI K.and KOGA T.: "Polypeptide compositions and antigenic homologies among plolamins from Italian, Common and Japanese millet cultivaars." J.Sci.Food Agric.72. 141-147 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] K.Takumi and T.Koga: "Frastiou and draroeterigaticu of Budiwbest proteins" Mum.Foc.Educ.Ehime Univ.Nat.Sci. 15. 99-109 (1994)

    • Related Report
      1994 Annual Research Report

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Published: 1994-03-31   Modified: 2016-04-21  

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