1993 年度実績報告書

生理機能の発現と組換え機構

研究課題

研究課題/領域番号	04262101
研究機関	東京大学
研究代表者	池田日出男東京大学, 医科学研究所, 教授 (10012775)
研究分担者	島田和典大阪大学, 微生物病研究所, 教授 (40037354) 駒野照弥東京都立大学, 理学部, 助教授 (00087131) 小林泰夫東京農工大学, 農学部, 教授 (10013319) 川市正史奈良先端科学技術大学院大学, 教授 (00195041) 大石道夫分子細胞生物学研究所, 教授 (00126004)
キーワード	免疫グロブリン遺伝子 / T細胞抗原レセプター遺伝子 / 部位特異的組換え / 非相同組換え / 標的遺伝子組換え / トランスポゾン / T-DNA / 遺伝子脆弱部位
研究概要	(1)細胞分化におけるDNAの再編成の機構。川市は、ショウジョウバエのRBP-Jκの遺伝が末梢神経系の発生に関与していることを、またマウスにおいては神経系を含む広範な組織の発生を調節する転写因子であることを示した。山岸は、マウスT細胞抗原レセプターα鎖J領域の上流から転写されるTEAを同定し、細胞分化の過程でα鎖遺伝子再配列に先行することを示した。大石は、従来のIGCR法に改良を加え、哺乳動物の1コピー/ゲノム以下のDNAの構造変化部位をクローニングする方法を確立し、酵母及びCHOゲノム中の特定遺伝子(ura及びAPRT)欠失部位の濃縮に成功した。(2)原核生物における遺伝子機能発現制御に関与する組換え機構。小林は、枯草菌のDNA再編成を引き起こす部位特異的組換え酵素の遺伝子spoIVCAが、胞子形成初期から中期の母細胞においてのみ機能するホロ酵素Eσ^Eによって転写されることを示した。駒野は、シャフロンのDNA再編成が、R64の接合伝達遺伝子群を強く発現している細胞では低く抑えられており、組換えが特殊な調節を受けていることを示唆した。(3)遺伝子の標的組み込みと非相同的組換え機構の解析。池田は、大腸菌のbio-λ領域における非相同組換えを解析する系を確立し、大腸菌recAとrecJ遺伝子の変異株において非相同組換えが減少することを見いだした。島田は、線状化したベクターDNAの両末端にヘアピン状のオリゴヌクレオタイドからなるキャップ構造を連結し、細胞内でのベクターの分解を防ぐことにより、標的遺伝子組換え体の選択の効率を飛躍的に上げる方法を確立した。町田は、AgrobacteriumのT-DNA、AcDNA転移因子、及び酵母の部位特異的組換え系を用いて植物遺伝子の突然変異を誘導する方法を開発した辻本は、遺伝性Fragile siteの分子構造を解明する目的で、第11染色体q13にマップされる遺伝性のFS(Frallq13.2)をモデルに選び、その脆弱部位のクローニングと構造解析を行った。

研究成果
(34件)

すべてその他

すべて文献書誌 (34件)

[文献書誌] Ikeda,H.: "Involvement of type II DNA topoisomerases in illegitimate recombination in Escherichia coli:Analyses of the partial diploid of phage T4 and the specialized transducing particles of phage λ." DNA topoisomerases in chemotherapy(Eds.T.Ando et al.). 49-56 (1993)
[文献書誌] Kato,J.: "Purification and characterization of DNA topoisomerase lV in Escherichia coli." DNA topoisomerases in chemotherapy(Eds.T.Ando et al.). 31-38 (1993)
[文献書誌] Kumagai,M.: "Effects of uvsX,uvxY and DNA topoisomerase genes on the formation of tandem duplications of the rII gene in bacteriophage T4." Genetics. 135. 255-264 (1993)
[文献書誌] Ikeda,H.: "DNA topoisomerase-mediated illegitimate recombination." DNA topoisomerases and their applications in pharmacology(Ed.L.F.Liu). (In press). (1994)
[文献書誌] Sugano,S.: "Molecular analysis of the dhp1^+ gene of Schizosaccharomyces pombe:an essential gene that has homology to the DST2 and RAT1 genes of Saccharomyces cerevisiae." Mol.Gen.Gen.(In press). (1994)
[文献書誌] Tatebayashi,K.: "Structural analyses of DNA fragments integrated by illegitimate recombination in Schizosaccharomyces pombe." Mol.Gen.Genet.(In press). (1994)
[文献書誌] Amakawa,R.: "Human Jκ recombination signal binding protein gene(IGKJPB):comparison with its murine homologue." Genomics. 17. 306-315 (1993)
[文献書誌] Shimizu,T.: "Mouse germline transcript of TCR alpha joining region and temporal expression in ontogeny." Int.Immunol.5. 155-160 (1993)
[文献書誌] Arakawa,H.: "The complete murine immunoglobulin class switch region of the alpha heavy chain gene-hierarchic repetitive structure and recombination breakpoints." J.Biol.Chem.268. 4651-4655 (1993)
[文献書誌] Kondo,T.: "Signal joint of immunoglobulin V lambda 1-J lambda and novel joints of chimeric V pseudogenes on extrachromosomal circular DNA from chicken bursa." Eur.J.Immunol.23. 245-249 (1993)
[文献書誌] Shimizu,T.: "Biased reading frames of pre-existing DH-JH coding joints and preferential nucleotide insertions at VH-DJH signal joints of excision products of immunoglobulin heavy chain gene rearrangements." EMBO J.11. 4869-4875 (1993)
[文献書誌] Goto,T.: "Projection structures of HIV-J observed with high resolution electron cryo-microscopy." J.Electron Microscopy. 43(In press). (1994)
[文献書誌] Iwasato,T.: "TGF-β induced class switch recombination may be modified during the cell proliferation restored by IL-2 stimulation." Immunol.Lett.(In press). (1994)
[文献書誌] Ogata,T.: "Chromosome 7 induces cellular senescence of non-tumorigenic immortalized human fibloblast cell lines." Mol.Cell.Biol.13. 6036 (1993)
[文献書誌] Kume,T.: "Induction of specific protein tyrosine phosphate transcripts at the stage of differentiation of mouse erthoroleukemia(MEL)cells." J.Biol.Chem.(In press). (1994)
[文献書誌] Yokota,H.: "A differetial cloning procedure of complex genomic DNA fragments." Analytical Biochem.(In press). (1994)
[文献書誌] Sasaki,Y.: "Construction of a normalized cDNA liblary by introduction of a semi-solid mRNA-cDNA hybridization system." Nucleic Acids Res.(In press). (1994)
[文献書誌] Kiyama,R.: "Instability of plasmid DNA maintenance caused by transcription of poly(dT)-containing sequences in E.coli:possible involvement of RNA DNA-hybrid triplexs formation." Nucleic Acids Res.(In press). (1994)
[文献書誌] Sato,J.: "Expression of the Bacillus subtilis spoIVCA gene,which encodes a site-specific recodmbinase,depends on the spoIIGB product." J.Bacteriol.(In press). (1994)
[文献書誌] Tojo,N.: "Cloning and nucleotide sequence of the Myxococcus xanthus lon gene:Indispensability of lon for vegetative growth." J.Bacteriol.175. 2271-2277 (1993)
[文献書誌] Tojo,N.: "The lonD gene is homologous to the lon gene encoding an ATP-dependent protease and is essential for the development of Myxococcus xanthus." J.Bacteriol.175. 4545-4549 (1993)
[文献書誌] Kim,S.-R.: "Nucleotide sequence and characterization of the traABCD region of IncII plasmid R64." J.Bacteriol.175. 5035-5042 (1993)
[文献書誌] V.Episkopou,S.: "Disruption of the transthyretin gene results in mice with depressed levels of plasma retinol and thyroid hormone." Proc.Natl.Acad.Sci.USA. 90. 2375-2379 (1993)
[文献書誌] Yamamura,K.: "Transgenic mouse model for human genetic diseases." Molec.Reproduc.Develop.36. 248-250 (1993)
[文献書誌] Matsuura,T.: "Four novel gene mutations in five Japanese male patients with neonatal or late onset OTC deficiency:Application of PCR single strand conformation polymorphisms for all exons and adjacent introns." Hum.Genet.(In press). (1993)
[文献書誌] Horie,K.: "Structure of replacement vectors for efficient gene targeting." J.Biochem.(Tokyo). (In press). (1994)
[文献書誌] Yoshida,Y.: "Molecular characterization of a short interspersed repetitive element from tabacco that exibits sequence homology to specific tRNAs." Proc.Natl.Acad.of Sci.USA. 15. 6562-6566 (1993)
[文献書誌] Banno,H.: "NPK1,a tabacco gene that encodes a protein with a domain homologous to yeast BCK1,STE11,and Byr2 protein kinases." Mol.Cell.Biol.13. 4745-4752 (1993)
[文献書誌] Akao,Y.: "Long-range mapping of the 11q23 region involved in chromosome aberrations in human tumors by pulse-field gel electrophoresis with a yeast artificial coromosome." Genes Chromo.Cancer. 8. 167-171 (1993)
[文献書誌] Tsujimoto,Y.: "Human B-cell neoplasia with the t(11;14)(q13;q32)and t(14;18)(q21;q32)chromosome translocations." The causes and consequences of chromosomal aberrations,ed.Kirsch,I.R.333-350 (1993)
[文献書誌] Dubois-Dauphin,M.: "Motoneurons overexpressing the bcl-2 protooncogene do not degenerate after axotomy in newborn transgenic mice." Proc.Natl.Acad.Sci.USA. (In press). (1994)
[文献書誌] Machida,Y.: "Molecular interactions between Agrobacterium and plants cells." The symposium book of 6th ISMP(ed.E.W.Nester). 85-96 (1993)
[文献書誌] Yoshioka,Y.: "Molecular characterization of a short interspersed repetitive elements from tobacco that exhibits sequence homology to specific tRNAs." Proc.Natl.Acad.Sci.USA. 90. 6562-6566 (1993)
[文献書誌] Shimoda,N.: "Genetic evidence for an interaction between the VirA sensor protein and the ChvE sugar-binding protein of Agrobacterium." J.Biol.Chem.268. 26552-26558 (1993)

1993 年度 実績報告書

生理機能の発現と組換え機構

研究代表者

池田 日出男 東京大学, 医科学研究所, 教授 (10012775)

研究成果

[文献書誌] Ikeda,H.: "Involvement of type II DNA topoisomerases in illegitimate recombination in Escherichia coli:Analyses of the partial diploid of phage T4 and the specialized transducing particles of phage λ." DNA topoisomerases in chemotherapy(Eds.T.Ando et al.). 49-56 (1993)

[文献書誌] Kato,J.: "Purification and characterization of DNA topoisomerase lV in Escherichia coli." DNA topoisomerases in chemotherapy(Eds.T.Ando et al.). 31-38 (1993)

[文献書誌] Kumagai,M.: "Effects of uvsX,uvxY and DNA topoisomerase genes on the formation of tandem duplications of the rII gene in bacteriophage T4." Genetics. 135. 255-264 (1993)

[文献書誌] Ikeda,H.: "DNA topoisomerase-mediated illegitimate recombination." DNA topoisomerases and their applications in pharmacology(Ed.L.F.Liu). (In press). (1994)

[文献書誌] Sugano,S.: "Molecular analysis of the dhp1^+ gene of Schizosaccharomyces pombe:an essential gene that has homology to the DST2 and RAT1 genes of Saccharomyces cerevisiae." Mol.Gen.Gen.(In press). (1994)

[文献書誌] Tatebayashi,K.: "Structural analyses of DNA fragments integrated by illegitimate recombination in Schizosaccharomyces pombe." Mol.Gen.Genet.(In press). (1994)

[文献書誌] Amakawa,R.: "Human Jκ recombination signal binding protein gene(IGKJPB):comparison with its murine homologue." Genomics. 17. 306-315 (1993)

[文献書誌] Shimizu,T.: "Mouse germline transcript of TCR alpha joining region and temporal expression in ontogeny." Int.Immunol.5. 155-160 (1993)

[文献書誌] Arakawa,H.: "The complete murine immunoglobulin class switch region of the alpha heavy chain gene-hierarchic repetitive structure and recombination breakpoints." J.Biol.Chem.268. 4651-4655 (1993)

[文献書誌] Kondo,T.: "Signal joint of immunoglobulin V lambda 1-J lambda and novel joints of chimeric V pseudogenes on extrachromosomal circular DNA from chicken bursa." Eur.J.Immunol.23. 245-249 (1993)

[文献書誌] Shimizu,T.: "Biased reading frames of pre-existing DH-JH coding joints and preferential nucleotide insertions at VH-DJH signal joints of excision products of immunoglobulin heavy chain gene rearrangements." EMBO J.11. 4869-4875 (1993)

[文献書誌] Goto,T.: "Projection structures of HIV-J observed with high resolution electron cryo-microscopy." J.Electron Microscopy. 43(In press). (1994)

[文献書誌] Iwasato,T.: "TGF-β induced class switch recombination may be modified during the cell proliferation restored by IL-2 stimulation." Immunol.Lett.(In press). (1994)

[文献書誌] Ogata,T.: "Chromosome 7 induces cellular senescence of non-tumorigenic immortalized human fibloblast cell lines." Mol.Cell.Biol.13. 6036 (1993)

[文献書誌] Kume,T.: "Induction of specific protein tyrosine phosphate transcripts at the stage of differentiation of mouse erthoroleukemia(MEL)cells." J.Biol.Chem.(In press). (1994)

[文献書誌] Yokota,H.: "A differetial cloning procedure of complex genomic DNA fragments." Analytical Biochem.(In press). (1994)

[文献書誌] Sasaki,Y.: "Construction of a normalized cDNA liblary by introduction of a semi-solid mRNA-cDNA hybridization system." Nucleic Acids Res.(In press). (1994)

[文献書誌] Kiyama,R.: "Instability of plasmid DNA maintenance caused by transcription of poly(dT)-containing sequences in E.coli:possible involvement of RNA DNA-hybrid triplexs formation." Nucleic Acids Res.(In press). (1994)

[文献書誌] Sato,J.: "Expression of the Bacillus subtilis spoIVCA gene,which encodes a site-specific recodmbinase,depends on the spoIIGB product." J.Bacteriol.(In press). (1994)

[文献書誌] Tojo,N.: "Cloning and nucleotide sequence of the Myxococcus xanthus lon gene:Indispensability of lon for vegetative growth." J.Bacteriol.175. 2271-2277 (1993)

[文献書誌] Tojo,N.: "The lonD gene is homologous to the lon gene encoding an ATP-dependent protease and is essential for the development of Myxococcus xanthus." J.Bacteriol.175. 4545-4549 (1993)

[文献書誌] Kim,S.-R.: "Nucleotide sequence and characterization of the traABCD region of IncII plasmid R64." J.Bacteriol.175. 5035-5042 (1993)

[文献書誌] V.Episkopou,S.: "Disruption of the transthyretin gene results in mice with depressed levels of plasma retinol and thyroid hormone." Proc.Natl.Acad.Sci.USA. 90. 2375-2379 (1993)

[文献書誌] Yamamura,K.: "Transgenic mouse model for human genetic diseases." Molec.Reproduc.Develop.36. 248-250 (1993)

[文献書誌] Matsuura,T.: "Four novel gene mutations in five Japanese male patients with neonatal or late onset OTC deficiency:Application of PCR single strand conformation polymorphisms for all exons and adjacent introns." Hum.Genet.(In press). (1993)

[文献書誌] Horie,K.: "Structure of replacement vectors for efficient gene targeting." J.Biochem.(Tokyo). (In press). (1994)

[文献書誌] Yoshida,Y.: "Molecular characterization of a short interspersed repetitive element from tabacco that exibits sequence homology to specific tRNAs." Proc.Natl.Acad.of Sci.USA. 15. 6562-6566 (1993)

[文献書誌] Banno,H.: "NPK1,a tabacco gene that encodes a protein with a domain homologous to yeast BCK1,STE11,and Byr2 protein kinases." Mol.Cell.Biol.13. 4745-4752 (1993)

[文献書誌] Akao,Y.: "Long-range mapping of the 11q23 region involved in chromosome aberrations in human tumors by pulse-field gel electrophoresis with a yeast artificial coromosome." Genes Chromo.Cancer. 8. 167-171 (1993)

[文献書誌] Tsujimoto,Y.: "Human B-cell neoplasia with the t(11;14)(q13;q32)and t(14;18)(q21;q32)chromosome translocations." The causes and consequences of chromosomal aberrations,ed.Kirsch,I.R.333-350 (1993)

[文献書誌] Dubois-Dauphin,M.: "Motoneurons overexpressing the bcl-2 protooncogene do not degenerate after axotomy in newborn transgenic mice." Proc.Natl.Acad.Sci.USA. (In press). (1994)

[文献書誌] Machida,Y.: "Molecular interactions between Agrobacterium and plants cells." The symposium book of 6th ISMP(ed.E.W.Nester). 85-96 (1993)

[文献書誌] Yoshioka,Y.: "Molecular characterization of a short interspersed repetitive elements from tobacco that exhibits sequence homology to specific tRNAs." Proc.Natl.Acad.Sci.USA. 90. 6562-6566 (1993)

[文献書誌] Shimoda,N.: "Genetic evidence for an interaction between the VirA sensor protein and the ChvE sugar-binding protein of Agrobacterium." J.Biol.Chem.268. 26552-26558 (1993)

1993 年度実績報告書

池田日出男東京大学, 医科学研究所, 教授 (10012775)