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Analysis of transfer genes in plasmid R64

Research Project

Project/Area Number 06454005
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 遺伝
Research InstitutionTOKYO METROPOLITAN UNIVERSITY

Principal Investigator

KOMANO Teruya  Faculty of Sience, TOKYO METROPOLITAN UNIVERSITY,Professor, 理学部, 教授 (00087131)

Co-Investigator(Kenkyū-buntansha) FURUYA Nobuhisa  Faculty of Sience, TOKYO METROPOLITAN UNIVERSITY,Assistant, 理学部, 助手 (50244413)
Project Period (FY) 1994 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1996: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1995: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1994: ¥3,000,000 (Direct Cost: ¥3,000,000)
KeywordsPlasmid R64 / Conjugation / Shufflon / Thin pilus
Research Abstract

Conjugal transfer is an important process to exchange genetic material among bacterial cells. Forty-nine genes were found in the 54-kb transfer region of plasmid R64. To test the requirement of these genes for R64 conjugation, deletion and insertion mutations were introduced into all genes. For R64 surface mating, 24 transfer genes were found to be essential, while for liquid mating, additional 12 genes were required. The pnd gene was responsible for plasmid stability. Many gene products were detected by the maxicell method. The 12 genes required for liquid mating were responsible for the formation of R64 thin pilus. The purified R64 thin pilus consists of the pils and pilV products. The pils product was first synthesized as 22-kDa protein, and then processed to 19-kDa mature pilin mediated by the pilU product. To reveal the relationship between structure and function of the pilS gene, many pils mutants were isolated. DNA rearrangement of the shufflon selects one of the seven pilV genes with different C-terminal segments and determined the recipient specificity in liquid mating. Lipopolysaccharides in the recipient cells are suggested to function as receptors for pilV proteins from mating experiments using various rfa mutants of E.coli k-12 and S.typhimurium LT2 as recipient strains. The rci gene encoding shufflon-specific recombinase was overxpressed and the Rci protein was purified. An in vitro recombination system was constructed using the purified Rci protein. A plasmid cyrrying two tandemly repeated oriT sequences was constructed. A recombination event to lose a DNA segment between two oriT sequences was observed after mobilization. A recombination depending nikAB genes was also observed in the donor cells.

Report

(4 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • 1994 Annual Research Report
  • Research Products

    (27 results)

All Other

All Publications (27 results)

  • [Publications] Furuya,N.and T.Komano: "Surface exclusion gene of lncl1 plasmid R64:Nucleotide sequence and analysis of deletion mutants" Plasmid. 32. 80-84 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Komano et al.: "Determination of recipient specificity by DNA rearrangement of shufflon in plasmid R64" Journal of Molecular Biology. 243. 6-9 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Furuya,N.and T.Komano: "Specific binding of NikA protein to one arm of 17-base pair inverted repeat sequence within oriT of plasmid R64" Journal of Bacteriology. 177. 46-51 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Komano et al.: "Mating variation by DNA inversions of shufflon in plasmid R64" Advances in Biophysics. 31. 181-193 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Furuya,N.and T.Komano: "Nucleotide sequence and characterization of the trbABC region of the IncI1 plasmid R64" Journal of Bacteriology. 178. 1491-1497 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] A.Gyohda et al.: "Analysis of DNA inversions in the shufflon of Plasmid R64" Journal of Bacteriology. 179. 1867-1871 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Furuya, N.and T.Komano: "Surface exclusion gene of lncl1 plasmid R64 : Nucleotide sequence and analysis of deletion mutants." Plasmid. 32. 80-84 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Komano, T., S.-R.Kim, T.Yoshida and T.Nisioka: "Determination of recipient specificity by DNA rearrangement of suhfflon in plasmid R64." J.Mol. Biol.243. 6-9 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Furuya, N.and T.Komano: "Specific binding of NikA protein to one arm of 17-base pair inverted repeat sequences within oriT of plasmid R64" J.Bacteriol.177. 46-51 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Komano, T., S.-R.Kim and T.Yoshida: "Mating variation by DNA inversions of shufflon in plasmid R64" Adv. Biophys.31. 181-193 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Furuya, N.and T.Komano: "Nucleotide sequence and characterization of the trbABC region of the lncl1 plasmid R64 : existence of the pnd gene for plasmid maintenance within the transfer region" J.Bacteriol.178. 1491-1497 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Gyohda, A., N.Funayama, and T.Komano: "Analysis of DNA inversions in the shufflon of Plasmid R64." J.Bacteriol.179. 1867-1871 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Furuya,N.and T.Komano: "Specific binding of NikA protein to one arm of 17-base pair inverted repeat sequence within oriT of plasmid R64" Journal of Bacteriology. 177. 46-51 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Komana et al.: "Mating variation by DNA inversions of shufflon in plasmid R64" Advances in Biophysics. 31. 181-193 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Furuya,N.and T.Komano: "Nucleotide sequence and characterization of the trbABC region of the IncI1 plasmid R64." Journal of Bacteriology. 178. 1491-1497 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] N.Tojo et al.: "Integration of bacteriophage Mx8 into the Myxococcus xanthus chromosome causes a structural alteration at the C-terminal region of the IntP protein." Journal of Bacteriology. 178. 4004-4011 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] M.Ogawa et al.: "FruA,a putative transcription factor essential for the development of Myxococcus xanthus." Molecular Microbiology. 22. 757-767 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] A.Gyohda et al.: "Analysis of DNA inversions in the shufflon of Plasmid R64." Journal of Bacteriology. 179(in press). (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Furuya,N.and T.Komano: "Surface exclusion gene of incl1 prasmid R64:Nucleotide sequence and analysis of deletion mutants" Plasmid. 32. 80-84 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] T.Komano et al.: "Determination of recipient specificity by DNA rearrangement of shufflon in prasmid R64" Journal of Molecular Biology. 243. 6-9 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] Furuya,N.and T.Komano: "Specific binding of NikA protein to one arm of 17-base pair inverted repeat sequence within oriT of plasmid R64" Journal of Bacteriology. 177. 46-51 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] T.Komano et al.: "Mating variation by DNA inversions of shufflon in plasmid R64" Advances in Biophysics. 31. 181-193 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Furuya,N.and T.Komano: "Nucleotide sequence and characterization of the trbABC region of the IncI1 plasmid R64." Journal of Bacteriology. 178(in press). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] 古屋 伸久 等: "Surface exclusion gene of Incll plasmid R64:Nucleotide sequence and analysis of deletion mutants." Plasmid. 32. 80-84 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 駒野 照弥 等: "Determination of recipient specificity by DNA rearrangement of shufflon in plasmid R64." Journal of Molecular Biology. 243. 6-9 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 古屋 伸久 等: "Specific binding of NikA protein to one arm of 17-base pair inverted repeat sequences within oriT of plasmid R64." Journal of Bacteriology. 177. 46-51 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 駒野 照弥 等: "Mating variation by DNA inversions of shufflon in plasmid R64." Advances in Biophysics. 31. 181-193 (1995)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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