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Integration of Non-natural Amino Acids Into Protein

Research Project

Project/Area Number 04453161
Research Category

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

Allocation TypeSingle-year Grants
Research Field 物質生物化学
Research InstitutionTokyo Institute of Technology

Principal Investigator

NISHIKAWA Kazuya  Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology. Associate Professor, 生命理工学部, 助教授 (60109262)

Co-Investigator(Kenkyū-buntansha) ISHIKAWA Masahide  Interdisciplinary Graduate School of Science and Engineering, Tokyo Inst.Tech.,, 大学院総合理工学研究科, 助手 (50212858)
UEDA Takuya  Faculty of Engineering, The University of Tokyo, Assistant Professor, 工学部, 助手 (80184927)
Project Period (FY) 1992 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥6,200,000 (Direct Cost: ¥6,200,000)
Fiscal Year 1993: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1992: ¥4,200,000 (Direct Cost: ¥4,200,000)
KeywordsBiosynthesis of proteins / Non-natural amino acids / Microsurgery of RNA / Artificial proteins
Research Abstract

1) Several protecting groups of 5'-terminal and internucleotide phosphates and base amines were tested to obtain better yield of aminoacyl pCpCpA.Biotinyl-lysyl pCpA was designed and synthesized to make it easier to detect the amino acid after integration into proteins.
2) An efficient method for preparing tRNA which has an anticodon of arbitrary nucleotide sequence and lacks 3'-terminal pCpCpA,was developed by the use of mictosurgery technique and yeast tRNA^<Phe>, tRNA^<Tyr> and so on. Furthermore, a non-radioactive method for determining nucleotide sequences of low-molecular-weight RNA was developed.
3) Advantage in nuclease resistance and translational efficiency of phosphorothioate-containing mRNA (thio-mRNA) over natural mRNA was demonstrated in E.coli cell-free translation system. Effects of unusual polyamines on phenylalanyl-tRNA formation was investigated in cell-free systems of an extreme thermophile, Thermus themophilus HB8. Fundamental characteristics of cell-tree translation systems from yeast, Candida cylindracea and bovine mitochondriawere also examined.

Report

(3 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • Research Products

    (20 results)

All Other

All Publications (20 results)

  • [Publications] K.Ogawa,S.Takemura,and K.Nishikawa: "Anticodon Loop Structure of Torulopsis utilis tRNA^<Val> and Valine Acceptance." Journal of Advanced Science. 5. 43-49 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] I.Motoki,T.Yokogawa,T.Hasegawa,K.Watanabe and K.Nishikawa: "Higher-order Structure of Yeast tRNA^<Tyr> and Its Chimeric Mutants in Aqueous Solution As Studied by Hydroxyl Radical "Footprinting"." Journal of Advanced Science. 5. 85-90 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] H.Tohda,N.Chikazumi,T.Ueda,K.Nishikawa,and K.Watanabe: "Efficient expression of E. coli dihydrofolate reductase gene by an in vitro translation system using phosphorothioate mRNA." Journal of Biotechnology. 34. 61-69 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Uzawa,A.Yamagishi,K.Nishikawa and T.Oshima: "Effects of unusual polyamines on phenylalanyl-tRNA formation." Journal of Biochemistry. 115. 830-832 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Suzuki,T.Ueda,T.Yokogawa,K.Nishikawa and K.Watanabe: "Chracterization of serine and leucine tRNAs in an asporogenic yeast Candida cylindracea and evolutionary implications of genes for tRNA^<Ser>CAG responsible for translation of a non-universal genetic code." Nucleic Acids Research. 22. 115-123 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] C.Takemoto,T.Koike,T.Yokogawa,L.Benkowski,L.L.Spremulli,T.Ueda,K.Nishikawa and K.Watanabe: "The ability of bovine mitochondrial transfer RNA^<Met> to decode AUG and AUA codons." Biochimie. 77. 104-108 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Ogawa, S.Takemura, and K.Nishikawa: "Anticodon Loop Structure of Torulopsis utilis tRNA^<Val> and Valine Acceptance." J.Adv.Sci.5. 43-49 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] I.Motoki, T.Yokogawa, T.Hasegawa, K.Watanabe and K.Nishikawa: "Higher-order Structure of Yeast tRNA^<Tyr> and Its Chimeric Mutants in Aqueous Solution As Studied by Hydroxyl Radical "Footprinting"." J.Adv.Sci.5. 85-90 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] H.Tohda, N.Chikazumi, T.Ueda, K.Nishikawa, and K.Watanabe: "Effecient expression of E.coli dihydrofolate reductase gene by an in vitro translation system using phosphorothioate mRNA." J.Biotechnol.34. 61-69 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Uzawa, A.Yamagishi, K.Nishikawa and T.Oshima: "Effects of unusual polyamines on phenylalanyl-tRNA formation." J.Biochem.115. 830-832 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Suzuki, T.Ueda, T.Yokogawa, K.Nishikawa, and K.Watanabe: "Characterization of serine and leucine tRNAs in an asporogenic yeast Candida cylindracea and evolutionary implications of genes for tRNA^<Ser> CAG responsible for translation of a non-universal gentic code." Nucleic Acids Res.22. 115-123 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] C.Takemoto, T.Koike, T.Yokogawa, L.Benkowski, L.L.Spremulli, T.Ueda, K.Nishikawa and K.Watanabe: "The ability of bovine mitochondrial transfer RNA^<Met> to decode AUG and AUA codons." Biochimie. 77. 104-108 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Kazuko Ogawa: "Anticodon Loop Structure of Torulopsis utilis tRNA^<Val> and Valine Acceptance" J.Adv.Sci.5. 43-49 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Takahiro Matsuki: "Sequence Analysis of low-molecular-weight RNAs by the use of non-radioactive labeling" Nucleic Acids Symp.Ser.29. 75-76 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Hideki Tohda: "Efficient expression of E.coli dihydrofolate reductase gene by an in vitro expression system using phosphorothiorte mRNA." J.Biotechnol.(印刷中). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Ichiro Motoki: "Higher-order Stlucture of Yeast tRNA^<Tvr> and Its chimeric Mutants in Aqueous Solution As studied by Hydroxyl Radical“Foot printing"" J.Adv.Sci.(印刷中). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Tatsuya Niimi: "Recognition of the Auticodon Loop of tRNA^<Ile>_1 by Isoleucyl-tRNA Synthetase from Escherichia coli" Nucleosides and Nucleotides. (印刷中). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Norihiko Yamazaki: "Purification and characterization of tRNA(adenosine-1)-methyltransferase from Thermus thermophilus HB27" Nudeic Acids Symp.Ser.27. 141-142 (1922)

    • Related Report
      1992 Annual Research Report
  • [Publications] Hiroshi Ayukawa: "Alkylation reaction of phosphorus Dxyacids" Nucleic Acids Symp.ser.27. 91-92 (1922)

    • Related Report
      1992 Annual Research Report
  • [Publications] Shingo Makino: "Chemical synthesis of Cyfidine-5'-monophosphono-N-acetylheuramimic Acid(CMP-Neu5AC)" Tetrahedron Lett.

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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