1996 年度実績報告書

細菌の病原性の分子遺伝学的研究

研究課題

研究課題/領域番号	08307004
研究種目	基盤研究(A)
応募区分	総合
研究機関	山口大学
研究代表者	中澤晶子山口大学, 医学部, 教授 (40053053)
研究分担者	林英生筑波大学, 基礎医学系, 教授 (40033203) 笹川千尋東京大学, 医科学研究所, 教授 (70114494) 小熊恵二岡山大学, 医学部, 教授 (00002262) 檀原宏文北里大学, 薬学部, 教授 (40114558) 寺脇良郎信州大学, 医学部, 教授 (10014333)
キーワード	病原細菌 / 緑膿菌 / 赤痢菌Ipa蛋白 / P.gingivalis / 腸炎ビブリオ / B.fragilis / ボツリヌス毒素 / ヘリコバクター・ピロリ
研究概要	1.病原細菌の定着・増殖に関与する因子について以下の結果を得た。 (1)H.pyloriの尿素走化性は粘稠環境で亢進し、ウレアーゼ依存性である。 (2)縁膿菌のシデロフォア合成に関与するpvd領域は菌の生存に必須でない。 (3)腸炎ビブリオのfur遺伝子を単離し、その制御下にある遺伝子単離法を確立した。 (4)腸管凝集・粘着性大腸菌の粘着因子は16kDaの外膜蛋白である。 (5)P.gingivalisの線毛形成遺伝子はfimA領域外にも存在する。 2.病原細菌の細胞障害性因子について以下の結果を得た。 (1)黄色ブドウ球菌はアルカリショックに応答して数個の遺伝子を発現する。 (2)P.gingivalisのグリシルプロリルペプチダーゼを精製し、遺伝子の単離を開始した。 (3)B.fragilisのノイラミニダーゼ遺伝子nanHの上流の調節遺伝子nanRを同定した。 (4)C.histolyticumのコラゲナーゼ遺伝子を枯草菌に導入し本酵素精製法を確立した。 (5)ボツリヌス菌A型毒素前駆体のHAは毒素分子構成と細胞への結合・吸収に重要である。 (6)ブドウ球菌のロイコシジンとガンマヘモリジンは共通の毒素成分と固有の赤血球認識成分から構成される。 (7)緑膿菌の細胞溶解毒素は前駆体のC末が切断され活性化される。また、本酵素遺伝子を運搬するファージは緑膿菌に広く存在する。 (8)赤痢菌Ipa蛋白はα5β1インテグリンと結合し、菌の付着部位にインテグリンの集積とアクチンの重合を誘導し、細胞骨格蛋白の再構築を介して細胞内へ侵入する。 (9)サルモネラの病原遺伝子slyAはTTGを開始コドンとする。

研究成果

(27件)

すべてその他

すべて文献書誌 (27件)

[文献書誌] Abe M.et al.: "A genetic system of Burkholderia cepacia:construction of mobilizable transposons and a cloning vector." Gene. 174. 191-194 (1996)
[文献書誌] Abe M.et al.: "The dsbB Gene Product Is Required for Protease Production by Burkholderia cepacia." Infect.Immun.64(10). 4378-4380 (1996)
[文献書誌] Mizote T.et al.: "Urease-Independent Chemotactic Responses of Helicdobacter pylori to Urea,Urease Inhibitors,and Sodium Bicarbonate." Infect.Immun.65(4)in press. (1997)
[文献書誌] Mizote T.et al.: "The thiJ locus and its relation to phosphorylation of hydroxymethylpyrimidine in Escherichia coli." Microbiology(UK). 142. 2969-2974 (1996)
[文献書誌] Yamamoto T.et al.: "Detection of the enteroaggregative Escherichia coli heat-stable enterotoxin 1 gene sequences in enterotoxigenic E.coli stains phathogenic for humans." Infect.Immun.64. 1441-1445 (1996)
[文献書誌] Yamamoto T.et al.: "Characterization of a novel hemagglutinin of diarrhea-associated Escherichia coli that has characteristics of diffusely adhering E.coli and enteroaggregative E.coli" Infect.Immun.64. 3694-3702 (1996)
[文献書誌] Yamamoto T.et al.: "Detection and sequences of the enteroaggregative Escherichia coli heat-stable enterotoxin 1 gene in enterotoxigenic E.coli strains isolated from piglets and calves with diarrhea." J.Clin.Mircobiol.35. 223-227 (1997)
[文献書誌] Yamamoto T.et al.: "Comparison of the nucleotide sequence of enteroaggregative Escherichia coli heatstable enterotoxin 1 genes among diarrhea-associated Escherichia coli." FEMS Microbiol.Lett.147. 89-95 (1997)
[文献書誌] Nakayama K.et al.: "Involvement of Arginine-Specific Cysteine Protease(Arg-gingipain) in Fimbriation of Porphyromonas gingivalis." J.Bacteriol.178(10). 2818-2824 (1996)
[文献書誌] Watanabe K.et al.: "Sequence and Product Analyses of the Four Genes Downstream from the Fimbrilin Gene(fimA)of the Oral Amaerobe Porphyromonas gingivalis." Microbiol.Immunol.40(10). 725-734 (1996)
[文献書誌] Shimizu T.et al.: "Characterization of a toxin-deficient Clostridium perfringens strain KZ1340." Microbiol.Immunol.40. 141-145 (1996)
[文献書誌] Ohta T.et al.: "Atomic force microscopy proposes a nevel model for stem-loop structure thatbinds a heat shock protein in the Staphylococcus aureus HSP70 operon." Biochem.Biophys.Res.Comm.226. 730-734 (1996)
[文献書誌] Ba-thein,W.et al: "The virR/virS locus regulates the transcription of genes encoding extracellular toxin production in Clostridium perfringens." J.Bacteriol.178. 2514-2520 (1996)
[文献書誌] Ohtani K.et al.: "Collagenase gene(colA)is located in the 3′-flanking region of the perfringolysin O(pfoA)Iocus in Clostridium perfringens." FEMS Microbiol.Lett.146. 155-160 (1997)
[文献書誌] Kuwahara T.et al.: "Detection of Bacteroides fragilis by PCR assay targeting the neuraminidase-encoding gene." Letters in Applied Microbiology. 22. 361-365 (1996)
[文献書誌] Jung C.M.et al.: "Expression of the colH gene encoding Clostridium histolyticum collagenase in Bacillus subtilis and its application to enzyme purification." Microbiol.Immunol.40. 923-929 (1996)
[文献書誌] Moriishi K.et al.: "Mosaic structures of neurotoxins produced from Clostridium botulinum type C and D organisms." Biochim.Biophys.Acta.1307(2). 123-126 (1996)
[文献書誌] Inoue K.et al.: "Molecular composition of Clostridium botulinum type A progenitor toxins." Infect.Immun.64(5). 1589-1594 (1996)
[文献書誌] Moriishi K.et al.: "Molecular cloning of the gene encoding the mosaic neurotoxin composed of parts of botulinum neurotoxin types C1 and D and PCR detection of its gene from Clostridium botulinum type Corganisms." Appl.Environ. Microbiol.62(2). 662-667 (1996)
[文献書誌] Takeshi K.et al.: "Simple method for detection of Clostridium botulinum type A to F neurotoxin genes by polymerase chain reaction." Microbiol.Immnuol.40(1). 5-11 (1996)
[文献書誌] Kameyama K.et al.: "Analysis of the phospholipase C gene of Clostridium perfringens KZ 1340 isolated from Antarctic soil." Microbiol.Immunol.40(4). 255-263 (1996)
[文献書誌] Yamakawa K.et al.: "Enhancement of Clostridium difficle toxin production by biotin-limited conditions." J.Med. Microbiol.44(2). 111-114 (1996)
[文献書誌] MacSiomoin R.et al.: "Identification and characterization of ispA,a Shigella flexneri chromosomal gene essential for normal in vivo cell division and intercellular spreading." Mol.Microbiol.19. 599-609 (1996)
[文献書誌] Watarai M.et al.: "Interaction of Ipa proteins of Shigella flexneri with α5β1 integrin promotes entry of the bacteria into mammalian cells." J.Exp.Med.183. 991-999 (1996)
[文献書誌] Uchiya K.et al.: "Identification and charactrization of phoN-Sf,a gene on the large plasmid of Shigella flexneri 2a encodind a nonspecific phosphatase." J.Bacteriol.178. 4548-4554 (1996)
[文献書誌] Suzuki T.et al.: "Functional analysis of Shigella VirG domains essential for interaction with vinculin and actin-based motility." J.Biol.Chem.271. 21878-21885 (1996)
[文献書誌] Watarai M.et al.: "rho,a small GTP-binding protein, is essential for Shigella invasion of epithelial cells." J.Exp,Med.185. 281-292 (1997)

1996 年度 実績報告書

細菌の病原性の分子遺伝学的研究

研究代表者

中澤 晶子 山口大学, 医学部, 教授 (40053053)

研究成果

[文献書誌] Abe M.et al.: "A genetic system of Burkholderia cepacia:construction of mobilizable transposons and a cloning vector." Gene. 174. 191-194 (1996)

[文献書誌] Abe M.et al.: "The dsbB Gene Product Is Required for Protease Production by Burkholderia cepacia." Infect.Immun.64(10). 4378-4380 (1996)

[文献書誌] Mizote T.et al.: "Urease-Independent Chemotactic Responses of Helicdobacter pylori to Urea,Urease Inhibitors,and Sodium Bicarbonate." Infect.Immun.65(4)in press. (1997)

[文献書誌] Mizote T.et al.: "The thiJ locus and its relation to phosphorylation of hydroxymethylpyrimidine in Escherichia coli." Microbiology(UK). 142. 2969-2974 (1996)

[文献書誌] Yamamoto T.et al.: "Detection of the enteroaggregative Escherichia coli heat-stable enterotoxin 1 gene sequences in enterotoxigenic E.coli stains phathogenic for humans." Infect.Immun.64. 1441-1445 (1996)

[文献書誌] Yamamoto T.et al.: "Characterization of a novel hemagglutinin of diarrhea-associated Escherichia coli that has characteristics of diffusely adhering E.coli and enteroaggregative E.coli" Infect.Immun.64. 3694-3702 (1996)

[文献書誌] Yamamoto T.et al.: "Detection and sequences of the enteroaggregative Escherichia coli heat-stable enterotoxin 1 gene in enterotoxigenic E.coli strains isolated from piglets and calves with diarrhea." J.Clin.Mircobiol.35. 223-227 (1997)

[文献書誌] Yamamoto T.et al.: "Comparison of the nucleotide sequence of enteroaggregative Escherichia coli heatstable enterotoxin 1 genes among diarrhea-associated Escherichia coli." FEMS Microbiol.Lett.147. 89-95 (1997)

[文献書誌] Nakayama K.et al.: "Involvement of Arginine-Specific Cysteine Protease(Arg-gingipain) in Fimbriation of Porphyromonas gingivalis." J.Bacteriol.178(10). 2818-2824 (1996)

[文献書誌] Watanabe K.et al.: "Sequence and Product Analyses of the Four Genes Downstream from the Fimbrilin Gene(fimA)of the Oral Amaerobe Porphyromonas gingivalis." Microbiol.Immunol.40(10). 725-734 (1996)

[文献書誌] Shimizu T.et al.: "Characterization of a toxin-deficient Clostridium perfringens strain KZ1340." Microbiol.Immunol.40. 141-145 (1996)

[文献書誌] Ohta T.et al.: "Atomic force microscopy proposes a nevel model for stem-loop structure thatbinds a heat shock protein in the Staphylococcus aureus HSP70 operon." Biochem.Biophys.Res.Comm.226. 730-734 (1996)

[文献書誌] Ba-thein,W.et al: "The virR/virS locus regulates the transcription of genes encoding extracellular toxin production in Clostridium perfringens." J.Bacteriol.178. 2514-2520 (1996)

[文献書誌] Ohtani K.et al.: "Collagenase gene(colA)is located in the 3′-flanking region of the perfringolysin O(pfoA)Iocus in Clostridium perfringens." FEMS Microbiol.Lett.146. 155-160 (1997)

[文献書誌] Kuwahara T.et al.: "Detection of Bacteroides fragilis by PCR assay targeting the neuraminidase-encoding gene." Letters in Applied Microbiology. 22. 361-365 (1996)

[文献書誌] Jung C.M.et al.: "Expression of the colH gene encoding Clostridium histolyticum collagenase in Bacillus subtilis and its application to enzyme purification." Microbiol.Immunol.40. 923-929 (1996)

[文献書誌] Moriishi K.et al.: "Mosaic structures of neurotoxins produced from Clostridium botulinum type C and D organisms." Biochim.Biophys.Acta.1307(2). 123-126 (1996)

[文献書誌] Inoue K.et al.: "Molecular composition of Clostridium botulinum type A progenitor toxins." Infect.Immun.64(5). 1589-1594 (1996)

[文献書誌] Moriishi K.et al.: "Molecular cloning of the gene encoding the mosaic neurotoxin composed of parts of botulinum neurotoxin types C1 and D and PCR detection of its gene from Clostridium botulinum type Corganisms." Appl.Environ. Microbiol.62(2). 662-667 (1996)

[文献書誌] Takeshi K.et al.: "Simple method for detection of Clostridium botulinum type A to F neurotoxin genes by polymerase chain reaction." Microbiol.Immnuol.40(1). 5-11 (1996)

[文献書誌] Kameyama K.et al.: "Analysis of the phospholipase C gene of Clostridium perfringens KZ 1340 isolated from Antarctic soil." Microbiol.Immunol.40(4). 255-263 (1996)

[文献書誌] Yamakawa K.et al.: "Enhancement of Clostridium difficle toxin production by biotin-limited conditions." J.Med. Microbiol.44(2). 111-114 (1996)

[文献書誌] MacSiomoin R.et al.: "Identification and characterization of ispA,a Shigella flexneri chromosomal gene essential for normal in vivo cell division and intercellular spreading." Mol.Microbiol.19. 599-609 (1996)

[文献書誌] Watarai M.et al.: "Interaction of Ipa proteins of Shigella flexneri with α5β1 integrin promotes entry of the bacteria into mammalian cells." J.Exp.Med.183. 991-999 (1996)

[文献書誌] Uchiya K.et al.: "Identification and charactrization of phoN-Sf,a gene on the large plasmid of Shigella flexneri 2a encodind a nonspecific phosphatase." J.Bacteriol.178. 4548-4554 (1996)

[文献書誌] Suzuki T.et al.: "Functional analysis of Shigella VirG domains essential for interaction with vinculin and actin-based motility." J.Biol.Chem.271. 21878-21885 (1996)

[文献書誌] Watarai M.et al.: "rho,a small GTP-binding protein, is essential for Shigella invasion of epithelial cells." J.Exp,Med.185. 281-292 (1997)

1996 年度実績報告書

中澤晶子山口大学, 医学部, 教授 (40053053)