1989 Fiscal Year Annual Research Report

細菌病原性の分子遺伝学的研究

Research Project

Project/Area Number	62304036
Research Institution	The University of Tokyo
Principal Investigator	吉川昌之介東京大学, 医科学研究所, 教授 (80012714)
Co-Investigator(Kenkyū-buntansha)	山本達男順天堂大学, 医学部, 講師 (80095843) 寺脇良郎信州大学, 医学部, 教授 (10014333) 笹川千尋東京大学, 医科学研究所, 助教授 (70114494) 江崎孝行岐阜大学, 医学部, 講師 (90151977) 檀原宏文北里研究所, 室長 (40114558)
Keywords	病原細菌学 / 病原性 / ビルレンス遺伝子 / 組換えDNA実験 / 分子遺伝学的研究 / 遺伝子解析 / 病原菌 / プラスミド
Research Abstract	組換えDNA実験技術を始めとする分子遺伝学的技術を病原細菌の病原性の解析に応用するはことを目的としてこの総合研究班は結成された。研究分担者総数21名という大規模な班員構成からなり、各分担者の対象とする菌種も多技にわたり、ビルレンス遺伝子の存在部位も染色体性、プラスミド性、およびバクテリオファ-ジ性と異るため各分担者の研究達成の難易度には著しい差があった。組換え体の選択方法、汎用される宿主・ベクタ-系でクロ-ン化できるか否か、EK系を用いることができるか否か、仮にクロ-ン化できたとしてそれが完全に形質発現するかなどにも大きな差があった。この壁を乗り切るためにベクタ-系を開発するところから始めたり、新たに宿主に特殊の変異を生じさせたり、遺伝子導入のために特殊の方法を採用したり多くの試行錯誤が行われた。幸に過去2年間の長時間にわたる班会議の議論を通じてこれら問題点の解決のための示唆が与えられた。本年は更に議論を深めることによって比較的研究の進展のよくない、難しい問題の担当者の研究上の困難を克服させるため班会議を早期に開催した。その結果、各分担者それぞれがほぼ所期の目的を達した。セラチアの線毛、サルモネラの病原性、赤痢菌の病原性、大腸菌の表層構造、赤痢菌の抗原、らい菌の抗原、バクテロイデスの病原性、とくに線毛、腸管感染病原菌の粘着因子、コレラ菌の溶血毒、ナグビブリオの溶血毒、腸炎ビブリオの溶血毒、緑膿菌のサイトトキシン、Pseudomonas Cepaciaの溶血毒などにつき、その遺伝子の存在様式、遺伝的構造、塩基配列の決定、形質発現の調節機構、前駆体物質のプロセッシング機構などを明らかにし、その病原的意義の解明に一定の知見を得ることができた。

Research Products

(38 results)

All Other

All Publications (38 results)

[Publications] P.Pramoonjago: "Bactericidal activity of C9-deficient human serum"
[Publications] T.Ouo: "Cloning and expression of the Bactroides fragilis neuraminidase gene nanH in Eschenichia coli"
[Publications] P.Pramonjago: "Anticomplementary activity of Trat protein of Esherichia coli Rloo plasmid"
[Publications] H.Ibagon: "Detection of ST-type toxin genes in Klebsiella oxytoca and Citorobacter freundii with syntheort shire oligonucleotide probes."
[Publications] S.-L.Lin: "Isolation of rntraccllular multiplication mutants of Salmonella typhi"
[Publications] S.Akimoto: "Transformation of Bacteroides thetaiotaomicron by electroporation"
[Publications] T.Nakazawa: "Cloning and sequence analysis of the recA gene from Pseudomonas cepacia."
[Publications] T.Nakazawa: "Cloning and sequince analysis of the a hemolysin-like gene from Pseudomonas cepacia."
[Publications] M.Makino: "Construction of genomic library of Mycobacterium leprae gene and its expression in Streptomyces lividans" Internatl.J.Leprosy.
[Publications] T.Iida: "Cloning and expression of two genes encoding highly homologous hemolysins from a kanagawa phenomenon-positive vibrio para haemolyticus T4750 strain" Gene.
[Publications] T.Iida: "Cloning and expression of two genes encoding highly homologous hemolysins from a kanagawa phenomenon-positive vibrio para haemolyticus T4750 strain" Gene. P.Pramoonjago TraT.anticomplementary protein of Escherichia coli RIOO plasmid:monoclonal antibodies to TraT and purification of TraT protein:
[Publications] M.Abe: "Purification and properties of a heat-stable hemolysin from pseudomonas cepacia"
[Publications] Y.Yoshida: "Molecular cloning and characterization of formI antigen genes of shigella sonnei"
[Publications] Y.Takahashi: "Cloning and overproduction in Escherichia coli of the fimbrin gene and possible genes responsible for the fimbriation in an oral anaerobe Bacteroides gingivalis"
[Publications] S.Inouye: "Flagellation of pseudomoas putida is dependent on the ntra (rpon)gene." Mol.Gen.Genet.
[Publications] K.Kawahara: "Indentification and mapping of mba regions of the Selmonella choleraesais virulence plasmid PKDSC 5c responsible for the mouse bactirenia." Microbial Pathogenesis.
[Publications] M.Nishibuchi: "Comparative analysis of the hemolysin genes of vibrio cholerae non 01,V.minicus,and V.hollisae that are similar to the talh gene of V.parahaemolyticus." FEMS Microbiol.Letters.
[Publications] H.Taniguchi: "cloning and characterization of a gene encoding a new thermo-stable hemolysin from Vibrio Parahaemolyticus." FEMS Microbiol.Letters.
[Publications] K.Yamamoto: "Two step processing for actiration of cytolysin/hemolysin of vibrio cholerae 01,El Tor:nucleotide sequence of the structural gene(inlyA)and characterization of the processed products." Infect.Immun.
[Publications] N.Terakado: "Transposon-insertion mutogenesis of a genetic region encoding serum resistance in an 8okb plasmid of salmonella dublin" J.Gen.Microbiol.
[Publications] K.Kwahara: "Lipopolysaccharide alteration mediated by the virulence plasmid of salmonalla." Microbial pathogenesis. 7. 195-202 (1989)
[Publications] T.Sekizake: "Loss of virulence associated with plasmid curing of chicken pathogenic Escherichia coli." Jpn.J.Vet.Sci. 51. 659-661 (1989)
[Publications] T.Yamamoto.: "Vibrio choleral 01 adherence to human small intestinal M cells in Vitro." J.Infect.Dis.160. 168-169 (1989)
[Publications] M.Mori: "Indentification of species and capsular types of klebsiella clinical isolate with special reference to klobsiella planticola." Microbiol.Immuhol.33. 887-895 (1989)
[Publications] H.Watanabe: "Genetic amalysis of an invasion region by use of a Tn3-lactransposon and identification of a second positive regulator gene,invE,for ull invasion of shigella sonnei: significant homology of InvE with parB of pladmid p1." J.Bacteriol.(1990)
[Publications] N.Kido: "Detection of lipopolysaccharides by ethidium bromide staining agter sodium dodecyl sulfate polyscry lamide gel electrophoresis." J.Bacteriol.
[Publications] J.I.Kato: "Cloning of regions requived for contact hemolysis and entry into LLc-MK2 cells fran shigella sonnei form I plasmid.VirF is apositive rogulegtor gene for thses phenotypes." Infect.Immun. 57. 1391-1398 (1989)
[Publications] T.Moriya: "A cryptic fimbrial gene in serratia marcescens." J.Baeteriol.171. 6629-6636 (1989)
[Publications] N.Kido: "Partial deletion of the cloned rfb gene of Escherichia coli 09 results in synthesis of a new O-antigenic lipopolysaccharide" J.Bacteriol. 171. 3629-3633 (1989)
[Publications] H.Chosa: "Loss of viruleuce in shigella strains preserved in cultare collections due to molecular alferation of the invasion plasnid" Microbiol.Pathogenesis. 6. 337-342 (1989)
[Publications] K.Kawahara: "Lipoply saccharide alteration nediatid by the viruleuce plasmid of Salmenella." Microbiol Pathogenesis. 7. 195-202 (1989)
[Publications] T.Hayashi: "purification and charac terization of cytotoxin from the crued extract of Pseudomonas aeruginosa." Microbiol Pathogenesis.6. 103-112 (1989)
[Publications] Y.Ike: "Genetic analysis of the PAD1 hemolysin /bacteriocin determinant in Enterococcus.faecalis:Tn917 insertional mutagenesis and cloning" J.Bacteriol.172. 155-163 (1990)
[Publications] C.Sasakawa: "Functional organization and nucleotide sefuence of virulence Region-2 on the lorge virulence plasmid in shigella flexneri 2a." Molec.Microbiol. 3. 1191-1201 (1989)
[Publications] M.Yamada: "Molecular cloring and characterization of chromosomal viruleuce region KcpA of Shigella flexneri." Molec.Microbiol. 3. 207-213 (1989)
[Publications] B.Adler: "A dual transcriptional activation system for the 230 Kb plasmid gencs coding for viruleuce assoiated antigens of shigella flexneri" Molec.Microbiol. 3. 627-635 (1989)
[Publications] T.Hayashi: "Pseudomonas aeruginosa cytotoxin:the nucleotide sefueuce of the genes and the mechanism of activation of protoxin." Molec.Microbiol. 3. 861-868 (1989)
[Publications] M.C Lett: "virG,a plasmid-coded virulence gene of shigella flexnceri.identification of the virG protein and deternination of the complete coding segueucd" J.Baeteriol.171. 353-359 (1989)

1989 Fiscal Year Annual Research Report

細菌病原性の分子遺伝学的研究

Principal Investigator

吉川 昌之介 東京大学, 医科学研究所, 教授 (80012714)

Research Products

[Publications] P.Pramoonjago: "Bactericidal activity of C9-deficient human serum"

[Publications] T.Ouo: "Cloning and expression of the Bactroides fragilis neuraminidase gene nanH in Eschenichia coli"

[Publications] P.Pramonjago: "Anticomplementary activity of Trat protein of Esherichia coli Rloo plasmid"

[Publications] H.Ibagon: "Detection of ST-type toxin genes in Klebsiella oxytoca and Citorobacter freundii with syntheort shire oligonucleotide probes."

[Publications] S.-L.Lin: "Isolation of rntraccllular multiplication mutants of Salmonella typhi"

[Publications] S.Akimoto: "Transformation of Bacteroides thetaiotaomicron by electroporation"

[Publications] T.Nakazawa: "Cloning and sequence analysis of the recA gene from Pseudomonas cepacia."

[Publications] T.Nakazawa: "Cloning and sequince analysis of the a hemolysin-like gene from Pseudomonas cepacia."

[Publications] M.Makino: "Construction of genomic library of Mycobacterium leprae gene and its expression in Streptomyces lividans" Internatl.J.Leprosy.

[Publications] T.Iida: "Cloning and expression of two genes encoding highly homologous hemolysins from a kanagawa phenomenon-positive vibrio para haemolyticus T4750 strain" Gene.

[Publications] M.Abe: "Purification and properties of a heat-stable hemolysin from pseudomonas cepacia"

[Publications] Y.Yoshida: "Molecular cloning and characterization of formI antigen genes of shigella sonnei"

[Publications] Y.Takahashi: "Cloning and overproduction in Escherichia coli of the fimbrin gene and possible genes responsible for the fimbriation in an oral anaerobe Bacteroides gingivalis"

[Publications] S.Inouye: "Flagellation of pseudomoas putida is dependent on the ntra (rpon)gene." Mol.Gen.Genet.

[Publications] K.Kawahara: "Indentification and mapping of mba regions of the Selmonella choleraesais virulence plasmid PKDSC 5c responsible for the mouse bactirenia." Microbial Pathogenesis.

[Publications] M.Nishibuchi: "Comparative analysis of the hemolysin genes of vibrio cholerae non 01,V.minicus,and V.hollisae that are similar to the talh gene of V.parahaemolyticus." FEMS Microbiol.Letters.

[Publications] H.Taniguchi: "cloning and characterization of a gene encoding a new thermo-stable hemolysin from Vibrio Parahaemolyticus." FEMS Microbiol.Letters.

[Publications] K.Yamamoto: "Two step processing for actiration of cytolysin/hemolysin of vibrio cholerae 01,El Tor:nucleotide sequence of the structural gene(inlyA)and characterization of the processed products." Infect.Immun.

[Publications] N.Terakado: "Transposon-insertion mutogenesis of a genetic region encoding serum resistance in an 8okb plasmid of salmonella dublin" J.Gen.Microbiol.

[Publications] K.Kwahara: "Lipopolysaccharide alteration mediated by the virulence plasmid of salmonalla." Microbial pathogenesis. 7. 195-202 (1989)

[Publications] T.Sekizake: "Loss of virulence associated with plasmid curing of chicken pathogenic Escherichia coli." Jpn.J.Vet.Sci. 51. 659-661 (1989)

[Publications] T.Yamamoto.: "Vibrio choleral 01 adherence to human small intestinal M cells in Vitro." J.Infect.Dis.160. 168-169 (1989)

[Publications] M.Mori: "Indentification of species and capsular types of klebsiella clinical isolate with special reference to klobsiella planticola." Microbiol.Immuhol.33. 887-895 (1989)

[Publications] H.Watanabe: "Genetic amalysis of an invasion region by use of a Tn3-lactransposon and identification of a second positive regulator gene,invE,for ull invasion of shigella sonnei: significant homology of InvE with parB of pladmid p1." J.Bacteriol.(1990)

[Publications] N.Kido: "Detection of lipopolysaccharides by ethidium bromide staining agter sodium dodecyl sulfate polyscry lamide gel electrophoresis." J.Bacteriol.

[Publications] J.I.Kato: "Cloning of regions requived for contact hemolysis and entry into LLc-MK2 cells fran shigella sonnei form I plasmid.VirF is apositive rogulegtor gene for thses phenotypes." Infect.Immun. 57. 1391-1398 (1989)

[Publications] T.Moriya: "A cryptic fimbrial gene in serratia marcescens." J.Baeteriol.171. 6629-6636 (1989)

[Publications] N.Kido: "Partial deletion of the cloned rfb gene of Escherichia coli 09 results in synthesis of a new O-antigenic lipopolysaccharide" J.Bacteriol. 171. 3629-3633 (1989)

[Publications] H.Chosa: "Loss of viruleuce in shigella strains preserved in cultare collections due to molecular alferation of the invasion plasnid" Microbiol.Pathogenesis. 6. 337-342 (1989)

[Publications] K.Kawahara: "Lipoply saccharide alteration nediatid by the viruleuce plasmid of Salmenella." Microbiol Pathogenesis. 7. 195-202 (1989)

[Publications] T.Hayashi: "purification and charac terization of cytotoxin from the crued extract of Pseudomonas aeruginosa." Microbiol Pathogenesis.6. 103-112 (1989)

[Publications] Y.Ike: "Genetic analysis of the PAD1 hemolysin /bacteriocin determinant in Enterococcus.faecalis:Tn917 insertional mutagenesis and cloning" J.Bacteriol.172. 155-163 (1990)

[Publications] C.Sasakawa: "Functional organization and nucleotide sefuence of virulence Region-2 on the lorge virulence plasmid in shigella flexneri 2a." Molec.Microbiol. 3. 1191-1201 (1989)

[Publications] M.Yamada: "Molecular cloring and characterization of chromosomal viruleuce region KcpA of Shigella flexneri." Molec.Microbiol. 3. 207-213 (1989)

[Publications] B.Adler: "A dual transcriptional activation system for the 230 Kb plasmid gencs coding for viruleuce assoiated antigens of shigella flexneri" Molec.Microbiol. 3. 627-635 (1989)

[Publications] T.Hayashi: "Pseudomonas aeruginosa cytotoxin:the nucleotide sefueuce of the genes and the mechanism of activation of protoxin." Molec.Microbiol. 3. 861-868 (1989)

[Publications] M.C Lett: "virG,a plasmid-coded virulence gene of shigella flexnceri.identification of the virG protein and deternination of the complete coding segueucd" J.Baeteriol.171. 353-359 (1989)

吉川昌之介東京大学, 医科学研究所, 教授 (80012714)