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ノイラミニダーゼ遺伝子の導入による乳酸菌の機能改良に関する研究

Research Project

Project/Area Number 14770118
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Bacteriology (including Mycology)
Research InstitutionThe University of Tokushima

Principal Investigator

桑原 知巳  徳島大学, 大学院・ヘルスバイオサイエンス研究部, 助教授 (60263810)

Project Period (FY) 2002 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2004: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2003: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
Keywordsノイラミニダーゼ / 乳酸菌 / 免疫アジュバン / 腸管免疫 / ワクチン / 腸管感染症 / 遺伝子組換え / 粘膜IgA / 免疫アジュバンド / ノイラニニダーゼ / アジュバンド / IgA / コレラ毒素 / 腸内細菌
Research Abstract

Lactococcus lactis YIT2081株にClostridium perfringens, Salmonalla TypyhimuriumおよびBacteroides fragilis由来のノイラミニダーゼ遺伝子nanHを導入し、ノイラミニダーゼを産生する乳酸菌を作成した。これらのうち、最も培養上清中のノイラミニダーゼ活性が高かった菌株はC.perfringensのnanH遺伝子を導入したL.lactis YIT2081(pBCnanH1)であり、本菌株をマウスに経鼻投与すると抗原として用いた卵白アルブミンに対する糞便中の特異的IgA量がベクターコントロール群の3.4倍に上昇したことから、本菌株は粘膜アジュバントとして有用であると考えられた。しかしながら、経口接種した乳酸菌の腸管への定着率は極めて低いことが知られており、本年度はL.lactis YIT2081(pBCnanH1)の腸管への定着率を調べた。ノイラミニダーゼを産生するL.lactis YIT2081(pBCnanH1)と対照菌株であるL.lactis (pBE31)を10mlのMRS broth (Em 5ug/ml)で30℃16時間培養し、新鮮な5mlのMRS brothで再懸濁したものをそれぞれ7週齢の雄性ラット4匹に、1ml/匹の量を経口投与した。糞便は輸送用培地に採取したものを100倍と10,000倍に希釈し、TATAC(Em 20μg/ml)培地に塗布した。これを30℃で3日間培養し、コロニーを計数した。投与生菌数はL.lactis (pBE31)が1.0×10^9 CFU/ml、L.lactis YIT2081(pBCnanH1)が0.9×10^9 CFU/mlであった。各菌株の糞便中の生菌数測定の結果、両菌株とも投与8時間後までは全てのラットの糞便中から分離されたが、24時間以降では対照群の1個体にのみL.lactisが検出された。以上の結果より、ノイラミニダーゼ遺伝子の導入は、L.lactisの腸管への定着率に影響を及ぼさないことが明らかとなった。

Report

(3 results)
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (16 results)

All 2005 2004 Other

All Journal Article (6 results) Publications (10 results)

  • [Journal Article] Inhibitory effect of fluvastatin on ileal ulcer formation in rats induced by a nonsteroidal antiinflammatory drug.2005

    • Author(s)
      Hagiwara, M. et al.
    • Journal Title

      World J.Gastroenterol. 11

      Pages: 1040-1043

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Simultaneous detection of Bacteroides fragilis group species by leuB-directed PCR.2005

    • Author(s)
      Miki, T. et al.
    • Journal Title

      J.Med.Invest. 52

      Pages: 101-108

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Role of unbalanced growth of Gram-negative bacteria in ilea] ulcer formation in rats treated with a nonsteroidal anti-inflammatory drug.2004

    • Author(s)
      Hagiwara, M. et al.
    • Journal Title

      J.Med.Invest. 51

      Pages: 43-51

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Betulinic acid augments the inhibitory effects of vincristine on growth and lung metastasis of B16F1O melanoma cells in mice.2004

    • Author(s)
      Sawada, N. et al.
    • Journal Title

      Br.J.Cancer 90

      Pages: 1672-1678

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Genomic analysis of Bacteroides fragilis reveals extensive DNA inversions regulating cell surface adaptation.2004

    • Author(s)
      Kuwahara, T. et al.
    • Journal Title

      Proc.Natl.Acad.Sci.USA 101

      Pages: 14919-14924

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Inhibitory effects of Centella asiatica on azoxymethane-induced aberrant crypt focus formation and carcinogenesis in the intestines of F344 rats.2004

    • Author(s)
      Bunpo, P. et al.
    • Journal Title

      Food Chem.Toxicol. 42

      Pages: 1987-1997

    • Related Report
      2004 Annual Research Report
  • [Publications] M.Hagiwara, K.Kataoka, H.Arimochi, T.Kuwahara et al.: "Role of unbalanced growth of Gram-negative bacteria in ileal ulcer formation in rats treated with a nonsteroidal anti-inflammatory drug"J.Med.Invest.. 51. 43-51 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] S.Nakanishi, K.Kataoka, T.Kuwahara, Y.Ohnishi.: "Effects of high amylase maize starch and Clostridium butyricum on metabolism in colonic microbiota and formation of azoxymethane-induced aberrant crypt foci in the rat. colon."Microbiol.Immunol.. 47. 951-958 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Y.Ogata, M.Nishi, H.Nakayama, T.Kuwahara et al.: "Role of bile in intestinal barrier function and its inhibitory effect on bacterial translocation in obstructive jaundice in rats."J.Surg.Res.. 115. 18-23 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] H.Shibata, C.Shirakata, H.Kawasaki, Y.Sato, T.Kuwahara et al.: "Flavone markedly affects phenotypic expression of β-lactam resistance in methicillin-resistant Staphylococcus aureus strains isolated clinically."Biol.Pharm.Bull.. 26. 1478-1483 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] T.Ito, T.Kuwahara et al.: "Regional mutagenicity of heterocyclic amines in the intestine : mutation analysis of the cII gene in lambda/lacZ transgenic mice."Mutat.Res.. 539. 99-108 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] K.Kuroiwa, H.Nakayama, T.Kuwahara et al.: "Augmenting effect of acetic acid for acidification on bactericidal activity of hypochlorite solution."Lett.Appl.Microbiol.. 36. 46-49 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Kuroiwa., K., et al.: "Augmentig effect of acetic acid for acidification on bactericidal activity of hypochlorite solution"Lett. Appl. Microbiol.. 36. 46-49 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Narioka, J., et al.: "Promotion of bacterial translocation by major liver resection in obstructive jaundice in rats colonized predominantly with indigenous Escherichia coli"J. Med. Microbiol.. 51. 687-694 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kuwahara, T., et al.: "Physical and genetic map of the Bacteroides fragilis YCH46 chromosome"FEMS Microbiol. Lett.. 207. 193-197 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Koizumi, T., et al.: "Role of dietary phosphorus in the progression of renal failure"Biochem. Biophys. Res. Commun.. 295. 917-921 (2002)

    • Related Report
      2002 Annual Research Report

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

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