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Highly Efficient Production of Recombinant Gene by Repeated Fed-Batch Culture

Research Project

Project/Area Number 60470118
Research Category

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

Allocation TypeSingle-year Grants
Research Field 反応工学
Research InstitutionNagoya University

Principal Investigator

KOBAYASHI Takeshi  Nagoya University, 工学部, 教授 (10043324)

Co-Investigator(Kenkyū-buntansha) IIJIMA Shinji  Nagoya University, 工学部, 助教授 (00168056)
Project Period (FY) 1985 – 1986
Project Status Completed (Fiscal Year 1986)
Budget Amount *help
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1986: ¥1,800,000 (Direct Cost: ¥1,800,000)
KeywordsCopy number / Transcription efficiency / Plasmid / Stability / Nutrient concentration / Fed-batch culture / Tryptophan promoter / グルコースセンサー
Research Abstract

For overproduction of useful proteins, control of gene-copy number, stability of plasmid, control of transcription efficiency and control of nutrients are important in fed-batch culture of gene-engineered microorganisms. The recombinant E. coli harboring a runaway-replication plasmid was cultivated at 25゜C and then the temperature was shifted to 37゜C. The copy number increased drastically and the gene product was amplified almost 40-fold.
Effects of nutrients on <alpha> -amylase production and plasmid stability were studied in fed-batch culture of recombinant E. coli. Plasmid was maintained stably in a semi-synthetic medium but plasmid-free segregants were observed in a synthetic medium.
To control transcriptional efficiency, fundamental properties of the trp promoter were also investigated in fed-batch culture using a recombinant containing the lacZ gene controlled by this promoter. On-off regulation of the promoter was achieved by controlling the tryptophan level.

Report

(1 results)
  • 1986 Final Research Report Summary
  • Research Products

    (11 results)

All Other

All Publications (11 results)

  • [Publications] Shinji Iijima: J.Chem.Tech.Biotechnol.36(11). 539-546 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Shuji Kawai: J.Ferment.Technol.64(6). 503-510 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Satoru Mizutani: J.Chem.Eng.Japan. 19(2). 111-116 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Satoru Mizutani: J.Ferment.Technol.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Satoru Mizutani: Biotech.Progress.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Shinji Iijima: Appl.Microbiol.Biotechnol.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Shinji Iijima, Kenichi Kai, Satoru Mizutani and Takeshi Kobayashi: "Effects of Nutrients on <alpha> -Amylase Production and Plasmid Stability in Fed-Batch Culture of Recombinant Bacteria" J. Chem. Tech. Biotechnol.36(11). 539-546 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Shuji Kawai, Satoru Mizutani, Shinji Iijima and Takeshi Kobayashi: "On-Off Regulation of the Tryptophan Promoter in Fed-Batch Culture" J. Ferment. Technol.64(6). 503-510 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Satoru Mizutani, Shinji Iijima and Takeshi Kobayashi: "Fed-Batch Culture of Escherichia coli Harboring a Runaway-Replication Plasmid" J. Chem. Eng. Japan. 19(2). 111-116 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Satoru Mizutani, Shinji Iijima, Makoto Morikawa, Kazuyuki Shimizu, Masakazu Matsubara, Yasumasa Ogawa, Rokuro Izumi, Kuniko Matsumoto and Takeshi Kobayashi: "On-Line Control of Glucose Concentration Using an Automatic Glucose Analyzer" J. Ferment. Technol.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] Satoru Mizutani, Shinji Iijima, Kazuyuki Shimizu, Masakazu Matsubara, Kenji Soda and Takeshi Kobayashi: "Mathematical Modeling and Response Characteristics of Runaway Replication for Temperature Shift-up" Biotech. Progress.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary

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Published: 1987-03-31   Modified: 2016-04-21  

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