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Biochemical preparation of isotpe-labelled lipopolysaccharide and nmr analysis of host recognition mechanism of phage

Research Project

Project/Area Number 26660085
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Applied biochemistry
Research InstitutionMie University

Principal Investigator

Inagaki Minoru  三重大学, 生物資源学研究科, 教授 (20242935)

Research Collaborator KARITA Shuichi  三重大学, 大学院生物資源学研究科, 教授 (90233999)
TAKEDA Shigeki  群馬大学, 大学院理工学府, 教授 (80282854)
FUJITA Katsutoshi  独立行政法人・製品評価技術基盤機構NITE, 資源保存課主任
Project Period (FY) 2014-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywordsリポ多糖 / 糖転移酵素 / 相同組み換え / 変異株 / 糖鎖構造解析 / 電気泳動 / 質量分析 / R-コア糖鎖 / galE遺伝子 / 欠損株 / DOC-PAGE / LC-MS / φX174ファージ / 宿主認識 / 感染 / スパイクタンパク質 / UDPガラクトースエピメラーゼ / 大腸菌 / galE / waaクラスター / PCR
Outline of Final Research Achievements

For incorpolation of 13C isotope-labelled galactose into the non-reducing end of lipopolysaccharide core structure, galE gene on E. coli C bacterium was determined on the gene. The mutant of E. coli F2513 were prepared by deletion of Galactosyl transferase genes, waaW and waaT. However, those mutants doesn't propagete in the cultural medium, and not yet certified the altered saccharide stracture on R-core lipopolysaccharide. By using the ASKA(-) clone, JW3598AM, the protein fraction of WaaU (hetosyl transferase) was prepared and submitted to treat the Salmonella bacterium. The incorpolation of an unnatural Hep residue was confirmed on the lipopolysacchride of Salmonella, using DOC-PAGE and LC-ESI-MS.

Report

(4 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • 2014 Research-status Report
  • Research Products

    (1 results)

All 2016

All Presentation (1 results)

  • [Presentation] Positive correlation between inactivation and DNA release of bacteriophage phiX174 caused by LPSs of E. coli C and E. coli K-122016

    • Author(s)
      Hayata Yamagata, Minoru Inagaki
    • Organizer
      Phage Kenkyukai
    • Place of Presentation
      海洋研究開発機構(神奈川県・横須賀市)
    • Year and Date
      2016-10-21
    • Related Report
      2016 Annual Research Report

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Published: 2014-04-04   Modified: 2018-03-22  

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