細菌性病原因子の分子遺伝学的研究

1992 Fiscal Year Annual Research Report

• Project/Area Number: 03304030
• Research Institution: Shinshu University
• Principal Investigator: 寺脇 良郎 信州大学, 医学部, 教授 (10014333)
• Co-Investigator(Kenkyū-buntansha): 笹川 千尋 東大医科研, 助教授 (70114494) ; 池 康嘉 群馬大学, 医学部, 教授 (60125820) ; 大西 克成 徳島大学, 医学部, 教授 (10037400) ; 中澤 晶子 山口大学, 医学部, 教授 (40053053) ; 天児 和暢 九州大学, 医学部, 教授 (20078752)
• Keywords: 病原細菌 / 病原因子 / 病原遺伝子 / 調節遺伝子 / DNA塩基配列 / 病原性プラスミド / 病原性ファージ

Research Abstract

大多数の分担研究者は初年度にひき続きそれぞれ同一の病原因子に対する研究を深めたが、同一菌種の他の病原因子の検索へと進んだ者もある。全体的な成果は概ね次のようにまとめられる。
1.推定病原因子の病原性の確認:H.pyloriのウレアーゼ遺伝子を不活化しマウスに対する病原性の低下を確認した(中沢)。
2.新たな病原遺伝子の同定とクローン化:エルトールコレラ菌の新溶血素遺伝子(山本耕),鶏病大腸菌の自家凝集遺伝子(関崎),P.gingivalisの血球凝集素遺伝子(吉村)。
3.病原遺伝子の構造決定:黄色ブドウ球菌のロイコシジンとα溶血素遺伝子(神尾),ボツリヌスC型毒素とE型毒素遺伝子(小熊),腸チフス菌Vi抗原の合成と輸送に関する遺伝子群(江崎)。
4.病原因子発現に関与する調節因子の同定:緑膿菌アルカリプロテアーゼ産生に対する鉄イオンとエリスロマイシンの抑制換果(鬼田),D群赤痢菌の細胞侵入性遺伝子群の発現に及ぼすphの効果(渡辺)。
5.病原遺伝子に対する調節遺伝子の同立と機能:B群赤痢菌の細胞間拡散を支配するvirGを正に調節するvirKと、細胞侵入性遺伝子群およびvirGの発現を転写後正に調節するvacB遺伝子の解析(笹川),腸炎ビブリオの耐熱性遺伝子(溶血毒)発現を調節する遺伝子群(西渕),ウェルシュ菌の4毒素遺伝子に仂く調節遺伝子群(林),豚コレラ菌の菌血症を惹起する3遺伝子に対する正の調節遺伝子(檀原),セラチアの潜在性線毛遺伝子に対する調節遺伝子の存在とそれに与える温度効果(天児)。
6.その他:緑膿菌サイトトキシンプロファージの宿主菌染色体への組込み桟構の解明とサイトトキシン精製法の確立(寺脇),B.fragilisの染色体制限酵素地図の作成(大西),病原性大腸菌EAggEC診断用プローブの開発(山本達),腸球菌β溶血素・バクテリオシンの活性化桟構(池)。

Research Products

• [Publications] T.Hayashi: "Molecular analysis of a cytotoxin-converting phage,φCTX,of P.aeruginosa:structure of the attp-cos-ctx region and integration into serine-" Molec.Microbiol(in press). 7(5). (1993)
• [Publications] A.Rahman: "Molecular cloning and nucleotide sequence of leucocidin F-component hene(lukF)from methicillir resistant Staphylococcus aureus" Biochem.Biophys.Res.Commun.184. 640-646 (1992)
• [Publications] T.Sekizaki: "Acid-induced autoagglutination found in chicken pathogenic Escherichia coli strain" J.Vet.Med.Sci.54. 493-499 (1992)
• [Publications] T.Sekizaki: "Isolation and characterization of type 1 fimbriae from a chicken pathogenic Escherichia coli serotype O78" J.Vet.Med.Sci.54. 1145-1149 (1992)
• [Publications] Y.Ike: "Hyperhemolitic phenomena associated with insertions of Tn916 into the hemolysin determinant of the Enterococcus faecalis plasmid pAD1" J.Bacteriol. 174. 1801-1809 (1992)
• [Publications] Y.Ike: "Evidence that the hemolysin/bacteriocin phenotype of E.faecalis subsp.zymogenes can be determined by plasmids in different incompatibility group" J.Bacteriol.174. 8172-8177 (1992)
• [Publications] N.Nakata: "Identification and characterization of virK,a virulence associated large plasmid gene essential for intercellular spreading of S.flexneri" Mol.Microbiol.6. 2387-2395 (1992)
• [Publications] T.Tobe: "vacB,a novel chromosomal gene required for expression of virulence genes on the large plasmid of Shigella flexneri" J.Bacteriol.174. 6359-6367 (1992)
• [Publications] C.Sasakawa: "The large virulence plasmid of shigella" Cur.Top.Microbiol.Immunol.180. 21-44 (1992)
• [Publications] T.TAmamoto: "Drug resistance and adherence to human intestines of enterroaggregative Escherichia coli" J.Infect.Dis.165. 744-749 (1992)
• [Publications] M Uchimura: "Production of hemagglutinins and pili by Vibriomimicus and its adherence to human and rabbit small intestines in vitro" FEMS Microbiol.Lett.91. 73-78 (1992)
• [Publications] T.Yamamoto: "Localized,aggregative and diffuse adherence to HeLa cells,plastic and human small intestines by Escherichia coli isolated from patients wuth--" J.Infect.Dis.166. 1295-1310 (1992)
• [Publications] Qi Ming-Shan: "Virulence phenotypes of Shigella flexneri 2a avirulent mutant 2450 can be complemented by the plasmid-coded.positive regulator virF gene" FEMS Microbiol.Lett.92. 217-222 (1992)
• [Publications] H.Takeuchi: "Structural gene and complete amino acid sequence of Vibrio alginolyticus collagenase" Biochem.J.281. 703-708 (1992)
• [Publications] S.Kanamoto: "Site-directed mutagenesis of Glu-141 and His-223 in P.aeruginosa elastase:catalytic activity processing and protective activity against ---" Infect.Inmmun.61(4). (1993)
• [Publications] K.Watanabe: "Molecular cloning and expression of a major surface protein of prophylomonas gingivalis in Escherichia coli" FEMS Microbiol.Lett.42. 47-56 (1992)
• [Publications] E.Isogai: "Fimbriae from Prophylomonas gingivalis induce chemiluminescence response of macrophages in different manner to its lipopolysaccharide" Bifidobacteria Microflora. 11. 39-43 (1992)
• [Publications] K.Kishishita: "Sequence variation in the theromstable direct hemo-lysis-related hemolysin(trh)gene of Vibrio parahemolyticus" Appl.Environ.Microbiol.58. 2449-2457 (1992)
• [Publications] K.Baba: "Examination by site-directed mutagenesis of the amino acid residues of the thermostable direct hemolysin of Vibrio parahemolyticus requered for---" Mjicrobiol.Pathogen.12. 279-287 (1992)
• [Publications] M.Nishibuchi: "Enterotoxigenicity of V.parahemolyticus with and without genes encoding thermostable direct hemolysin" Infect.Inmmun.60. 3539-3545 (1992)
• [Publications] J.Tada: "Detection of the thermostable direct hemolysin gene(tdh)and the thermostable direct hemolysin related gene(tdh)of V.parahemolyticus by PCR" Mol.Cell.Probes. 6. 477-488 (1992)
• [Publications] S.Tamatsukuri: "Detection of a heat-labile entertoxin gene in enterotoxigeic Escherichia coli by densitometric evaluation using highly specific enzyme---" Eur.J.Clin.Microbiol.Inf.Dis.10. 1048-1055 (1991)
• [Publications] K.Yamamoto: "Enzyme-labeled oligonucleotide probes for detection of genes for thermostable direct hemolysin and TDH-related hemolysin of V.parahemolyticus" Can.J.Microbiol. 38. 410-413 (1992)
• [Publications] K.Miyagi: "Cholera diagnosed in clinical laboratory by hybridization" Lancet. 339. 988-989 (1992)
• [Publications] A.Naka: "Characterization of two forms of hemagglutinin/protease produced by Vibrio cholerae non-01" FEMS Microbiol.Lett.98. 197-200 (1992)
• [Publications] N.Fujii: "Cloning and whole mucleotide sequence of the gene for the light chain component of botulinum type E toxin from C.butyricum strain BL6430 and---" Microbiol.Inmmunol.36. 213-220 (1992)
• [Publications] K.Tsuzuki: "The complete nucleotide sequence of the gene coding for the nontoxic nonhemagglutinin component of C.botulinum type C progenitor toxin" Biochem.Biophys.Res.Commun.183. 1273-1279 (1992)
• [Publications] N.Yokosawa: "Inhibition of nonepinephrine secretion from dihitonin permialized PC12 cells by botulinum type D toxin" Toxicon. 30. 1555-1562 (1992)
• [Publications] H.Konishi: "Cytopathic effects of Helicobacter pylori on cultured mammalian cells" J.Med.Microbiol.37. 118-122 (1992)
• [Publications] M.Karita: "Establishment of new small animal model for human gastric Helicobacter pylori infection" Eur.J.Gastroenterol.Hepatol.Suppl.4. S25->29 (1992)
• [Publications] M.Tsuda: "Genetic transformation in Helicobacter pylori" Microbiol.Inmmunnol.37. (1992)
• [Publications] Y.Mizunoe: "Identification and nucletode sequence of the gene determining the adhesin capacity of Serratia marcescense" J.Bacteriol.173. 3257-3260 (1991)
• [Publications] H.Danbara: "Effect of 50 kb-plasmid,pKDSC50,of Salmonella choleraesuis RF-1 strin on pig septisemia" J.Vet.Med.Sci.54. 1175-1178 (1992)
• [Publications] M.Emoto: "Induction of gamma/delta T-cell in murine salmonellosis by an avirulent but not by a vilurent strain of Salmonella choleraesuis" J.Exp.Med.176. 363-372 (1992)
• [Publications] H.Matsui: "Molecular mechanism of regulation of expressior of plasmid-encoded mouse bacteremia(mba) genes in Salmonella serovar choleraesuis" Mol.Gen.Genet.54. 219-216 (1993)
• [Publications] P.A.Gulig: "Molecular analysis of spv virulence genes of the Salmonella virulence plasmids" Mol.Microbiol.(1993)
• [Publications] 秋本 茂: "ノイラミニダーゼ遺伝子を利用したBacteroides tragilis診断用プローブの開発" 日本臨床1992年特別号感染症-遺伝子診断と分子疫学. 642. (1992)