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A Trial for a Creation of a New Protein Fiber by Modification of Fibroin Gene, Bombyx mori L.

Research Project

Project/Area Number 62480046
Research Category

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

Allocation TypeSingle-year Grants
Research Field 蚕糸学
Research InstitutionUniversity of Osaka Prefecture (1988)
Tokyo University of Agriculture and Technology (1987)

Principal Investigator

HIMENO Michio  University of Osaka Prefecture, College of Agriculture, 農学部, 教授 (10026411)

Co-Investigator(Kenkyū-buntansha) WADANO Akira  University of Osaka Prefecture, College of Agriculture, 農学部, 講師 (40081575)
SHIGEMATU Masanori  Tokyo University of Agriculture and Technology,Faculty of Technol, 工学部, 助手 (90015032)
Project Period (FY) 1987 – 1988
Project Status Completed (Fiscal Year 1988)
Budget Amount *help
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1988: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1987: ¥4,900,000 (Direct Cost: ¥4,900,000)
KeywordsFibroin / Fibroin Gene / fibroin Crystalline Fraction / Protein of Silk / Protein Engineering / Fussed Protein / フィブロイン / フィブロイン遺伝子 / フィブロイン結晶領域 / ベッター作成 / 遺伝子改変
Research Abstract

The silk fibroin is excellent materials as a fiber of cloth and it is expected as good materials for medical instrument and salso for immobilization of enzyme. This project is a trial for a ceration of new protein fiber by modification of fibroin gene, Bombyx mori L..
Parts of a typical repeating amino acid sequence (- Ser - Gly - Ala - Gly - Ala - Gly -) in silk fibroin molecule are called a fibroin crystalline fraction (Fcp). If a radical amino acid be added in Fcp sequence, the character of fibroin protein will drastically changed. then in this project we attempt addition of the code of lysine and/or methionine in DNA of the Fcp sequence and expression of fibroin gene in escherichia coli.
First, a expression vector of Fcp DNA sequence was made. When a passenger DNA sequence of 3n + 1 base pair is introduced into poly linker site of - galactosidase ( -gal) gene on plasmid p41 (made in this project) carring tac promoter. the introduced dna sequence will be express as a fussed protein with -gal in E.coli. Next, the Fcp sequence in plasmid pBmF6 harboring the fibroin gene (kindlly provided by Prof. Y. Suzuki) was obtained by Pst I, Hae II and Bal 31 treatment of the plasmid, and also by ALu I partially treatment, then we got plasmid pFCP and pFKB. Third, four DNA sequences were synthesized by a DNA synthesizer. The fragment were linked to p41 and also mixture of the fragments and polymer of one were introduced the plasmid, then plasmid pFMS was obtained. The pFCP, pFKB pr pFMS DNA easily deleted in E. coli. When E. coli JM109 carring pFCP, pFKB or pFMS was cultured on a plate containing IPTG and X-gal, the colony become blue. The blue colonies also reacted anti-Fcp rat serum. The protein extract isolated the blue colonies were applied on page, and determined by western blotting method using anti-Fcp rat serum and anti-rat IgG-HRPO conjugated rabbit IgG. The blue colonies produced a Fcp like protein but the molecular weight were not determined.

Report

(3 results)
  • 1988 Annual Research Report   Final Research Report Summary
  • 1987 Annual Research Report
  • Research Products

    (3 results)

All Other

All Publications (3 results)

  • [Publications] 姫野道夫,神阪敏行,深田哲夫,富畑賢司,丸山真樹子,小松原秀介: 日本農芸化学会誌. 63. 122-123 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M. Himeno; T. Kohosaka; T. Fukada; K. Tomihata; M. Maruyama; H. Komatsubara: "Expression of Sythesized Model Fibroin DNA in Escherichia coli." Nippon Nogeikagaku Kaishi. 63. 122-123 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 姫野道夫、神阪敏行、深田哲夫、富畑賢司、丸山真樹子、小松原秀介: 日本農芸化学会誌. 63. 122-123 (1989)

    • Related Report
      1988 Annual Research Report

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Published: 1987-04-01   Modified: 2016-04-21  

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