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Development of in vitro DNA virus autonomously evolving in vitro

Research Project

Project/Area Number 14580666
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionSaitama University, Faculty of Engineering

Principal Investigator

HUSIMI Yuzuru  Saitama University, Dept of Functional Materials Science, Professor, 工学部, 教授 (80011641)

Project Period (FY) 2002 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2003: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2002: ¥2,000,000 (Direct Cost: ¥2,000,000)
KeywordsEvolutionary molecular engineering / Genotype-phenotype Linking / in vitro virus / Artificial Life / Random peptide library / Isothermal DNA amplification / Genetic information / Evolution reactor / 遺伝子型表型対応付け / mRNA提示法 / T7プロモータ
Research Abstract

We established an isothermal amplification protocol for DNA/RNA at 50℃ using a thermostable T7 RNA polymerase and 1.4M trehalose. Using this protocol, a natural selection-type evolution reactor for DNA having a {A, T, G} random region was driven. Selected sequences were apt to be AT-rich. Evolution of amplification mechanism from 3SR-scheme to RNA-Z scheme was not observed. The optimum T7 promoter sequence at 50℃ was at the point of Hamming distance 2 from that at 37℃. We also confirmed the quality of DNA libraries for random peptide library of the controlled amino acid composition developed by us (MLSDS method). We studied the evolution dynamics in the evolution reactors using various fitness landscape model. Introducing the evolutionary temperature as a function of mutation rate and population size, we were able to define the free fitness as a Lyapunov function of this process. We could not succeed to integrate all the results into an autonomously evolving molecular entity.

Report

(3 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • Research Products

    (35 results)

All Other

All Publications (35 results)

  • [Publications] Aita T.: "Surveying a Local Fitness Landscape of a Protein with Epistatic Sites for the study of Directed Evolution"Biopolymers. 64. 95-105 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Husimi Y.: "Correlated Flexible Molecular Coding and Molecular Evolvability"J.Biol.Phys.. 28. 499-507 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kitamura K.: "Construction of block shuffled libraries of DNA for evolutionary protein engineering : Y-ligation based block shuffling"Protein Engineering. 15. 843-853 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tabuchi I.: "An efficient ligation in the making of in vitro virus for in vitro protein evolution"Biological Proced.Online. 4. 49-54 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Aita T.: "Statistical Formulae for Energy Distribution among a Globular Protein Structure Ensemble"J.Theor.Biol.. 220. 107-121 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Aita T.: "An in silico exploration of Neutral network in protein sequence space"J.Theor.Biol.. 221. 599-613 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Aita T.: "Thermodynamical Interpretation of Adaptive Walk on a Mt.Fuji-type Fitness Landscape : Einstein's Relation-like Formula holds in a Stochastic Evolution"J.Theor.Biol.. 225. 215-228 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Suzuki M.: "Intramolecular Fluorescent Resonance Energy Transfer (FRET) by BODIPY Chemical Modification of Cysteine-engineered Mutants of Green Fluorescent Protein"Chem.Lett.. 32. 306-307 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Aita T.: "Thermodynamic Interpretation of evolutionary dynamics on a Mt.Fuji-type fitness landscape"Bulletin Math.Biology. In press. (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 伏見 譲: "In vitro virusと進化分子化学"蛋白質・核酸・酵素. 48. 1481-1487 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 伏見 譲(編集および分担): "シリーズ・ニューバイオフィジックスII第8巻「生命の起源と進化の物理学」"共立出版. 231 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 伏見 譲(編集および分担): "生命の起源-「物質の進化」から「生命の進化」へ"丸善. 138 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Aita T., Hamamatsu N., Nomiya Y, Uchiyama H., Shibanaka Y., Husimi Y.: "Surveying a Local Fitness Landscape of a Protein with Epistatic Sites for the study of Directed Evolution."Biopolymers. 64. 95-105 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Husimi Y., Aita T., Tabuchi I.: "Correlated Flexible Molecular Coding and Molecular Evolvability."J.Biol.Phys.. 28. 499-507 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kitamura K., Kinoshita Y., Narazaki S., Nemoto N., Husimi Y., Nishigaki K.: "Construction of block shuffled libraries of DNA for evolutionary protein engineering : Y-ligation based block shuffling."Protein Engineering. 15. 843-853 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tabuchi I., Soramoto S., Suzuki M., Nemoto N., Husimi Y.: "An efficient ligation in the making of in vitro virus for in vitro protein evolution."Biological Proced.Online. 4. 49-54 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Aita T., Husimi Y.: "Statistical Formulae for Energy Distribution among a Globular Protein Structure Ensemble."J.Theor.Biol.. 220. 107-121 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Aita T., Ota M., Husimi Y.: "An in silico exploration of Neutral network in protein sequence space."J.Theor.Biol.. 221. 599-613 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Aita T., Husimi Y.: "Thermodynamical Interpretation of Adaptive Walk on a Mt.Fuji-type Fitness Landscape : Einstein's Relation-like Formula holds in a Stochastic Evolution."J.Theor.Biol.. 225. 215-228 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Suzuki, M., et al.: "Intramolecular Fluorescent Resonance Energy Transfer (FRET) by BODIPY Chemical Modification of Cysteine-engineered Mutants of Green Fluorescent Protein."Chem.Lett.. 32. 306-307 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Aita, T., Husimi, Y.: "Thermodynamic Interpretation of evolutionary dynamics on a fitness landscape in an evolution reactor I."Bulletin Math.Biology. (in press). (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Husimi Y., Tabuchi I.: "In vitro virus and Evolutionary Molecular Engineering"Protein, NucleicAcid, Enzyme. 4. 1481-1487 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Aita T.: "An in silico exploration Neutral network in protein sequence space."J.Theor.Biol. 221. 599-613 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Aita T.: "Thermodynamical Interpretation of Adaptive Walk on a Mt.Fuji-type Fitness Landscape : Einstein's Relation-like Formula holds in a Stochastic Evolution"J.Theor.Biol.. 225. 215-228 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Suzuki M.: "Intramolecular Fluorescent Resonance Energy Transfer (FRET) by BODIPY Chemical Modification of Cysteine-engineered Mutants of Green Fluorescent Protein."Chem.Lett.. 32. 306-307 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Aita T.: "Thermodynamic Interpretation of evolutionary dynamics on a Mt.Fuji-type fitness landscape."Bulletin Math.Biology. (in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] 伏見 譲: "In vitro virusと進化分子工学"蛋白質・核酸・酵素. 48. 1481-1487 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 伏見 譲(編集および分担): "生命の期限-「物質の進化」から「生命の進化」へ"丸善. (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Aita T.: "Surveying a Local Fitness Landscape of a Protein with Epistatic Sites for the study of Directed Evolution"Biopolymers. 64. 95-105 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Husimi Y.: "Correlated Flexible Molecular Coding and Molecular Evolvability"J. Biol. Phys.. 28. 499-507 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kitamura K.: "Construction of block shuffled libraries of DNA for evolutionary protein engineering : Y-ligation based block shuffling"Protein Engineering. 15. 843-853 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Tabuchi I.: "An efficient ligation in the making of in vitro virus for in vitro protein evolution"Biological Proced. Online. 4. 49-54 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Aita T.: "Statistical Formulae for Energy Distribution among a Globular Protein Structure Ensemble"J. Theor. Biol.. 220. 107-121 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Aita T.: "An in silico exploration of Neutral network in protein sequence space"J. Theor. Biol.. 221(in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 伏見 譲(編集および分担): "シリーズ・ニューバイオフィジックスII第8巻「生命の起源と進化の物理学」"共立出版. 231 (2003)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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