2003 Fiscal Year Annual Research Report

利己的な制限酵素修飾酵素遺伝子のゲノム進化での役割に関する比較ゲノムと実験進化

Research Project

Project/Area Number	15370099
Research Institution	The University of Tokyo
Principal Investigator	小林一三東京大学, 医科学研究所, 助教授 (30126057)
Keywords	ゲノム / 最近 / バイオテクノロジー / 制限酵素 / 病原細菌 / ゲノム進化 / 利己的遺伝子 / ゲノム再編
Research Abstract	(1)ゲノム情報からの解析。 (1-1)解読ゲノムからの制限修飾遺伝子の探索。ある解読好熱古細菌ゲノム中に、DNAメチル化酵素ホモログを洗い出し、その近隣のORFについて、制限酵素遺伝子候補を絞り、これら候補ORFをPCRでふやし、小麦胚芽のin vitroたんぱく合成系で発現させ、DNA切断活性を検索した。制限酵素を発見し、その反応条件、認識配列、切断位置を決定した。これは、新規の断端をもっていた。ほぼ完全に精製した。 (1-2)ゲノム多型形成の痕跡の探索。 Neisseria meningitides、Staphylococcus aureusについて近縁の細菌の全ゲノム配列を比較し、新しいゲノム多型の形成機構を発見した。 (2)制限酵素修飾酵素遺伝子の自己増殖とゲノム再編。 (2-1)同調的自己増幅誘発。 RM遺伝子が増幅したか否かを検討するためのドットブロットによる簡便なアッセイ系を作った。 (2-2)制限修飾遺伝子のサブゲノム間転移。大腸菌内での制限修飾酵素遺伝子のプラスミドから染色体への転移が制限酵素活性に依存するかを検討し、結論を得た。 (2-3)大規模ゲノム再編。染色体上にPaeR71制限修飾遺伝子と相同なゲノム領域をタンデムに持つ大腸菌を繰り返し植え継ぎ、どのようなゲノムが生き残るかを調べた。 (2-4)非染色体DNAに対する制限酵素の攻撃と再編。制限修飾遺伝子の攻撃によってドライブされるDNAの再編を、プラスミドで解析した。

Research Products

(15 results)

All Other

All Publications (15 results)

[Publications] A.Ino, S.Yamamoto, Y.Kaneda, I.Kobayashi: "Somatic gene targeting with RNA/DNA chimeric oligonucleotides : an analysis with a sensitive reporter mouse system."J.Gene Medicine. (刊行中).
[Publications] Y.Fujitani, I.Kobayashi: "Random-walkers active in genetic recombination."Biology International. (刊行中).
[Publications] K.Kusano, Y.Asami, A.Fujita, M.Tanokura, I.Kobayashi: "Type I restriction enzyme with RecA protein promotes illegitimate recombination."Plasmid. 50. 202-212 (2003)
[Publications] N.Handa, I.Kobayashi: "Accumulation of large non-circular forms of the chromosome in recombination-defective mutants of Escherichia coli."BMC Molecular Biology. 4. 5 (2003)
[Publications] Y.Katayama, F.Takeuchi, T.Ito, X.X.Ma, Y.Mizutani-Ui, I.Kobayashi, K.Hiramatsu: "Identification in a Methicillin-Susceptible Staphylococcus hominis of an Active Primordial Mobile Genetic Element for the Staphylococcal Cassette Chromosome mec (SCCmec) which - in Methicillin-Resisitant Staphylococcus aureus - is the Carrier of the Resisitance Gene mec."J.Bacteriol.. 185. 2711-2722 (2003)
[Publications] R.J.Roberts, et al.: "A nomenclature for restriction enzymes, DNA methyltransferases, homing endonucleases and their genes."Nucleic Acids Res. 31. 1805-1812 (2003)
[Publications] M.Sadykov, Y.Asami, H.Niki, N.Handa, M.Itaya, M.Tanokura, I.Kobayashi: "Multiplication of a restriction modification gene complex."Mol.Microbiol. 48. 417-427 (2003)
[Publications] N.Takahashi, H.Yoshikura, I.Kobayashi: "An Escherichia coli strain, BJ5183, that shows highly efficient conservative (two-progeny) DNA double-strand break repair of restriction breaks."Gene. 303. 89-97 (2003)
[Publications] Y.Fujitani, I.Kobayashi: "Asymmetric Random Walk in a Reaction Intermediate of Homologous Recombination."J.Theor.Biol.. 220. 359-370 (2003)
[Publications] 小林一三, 石川健: "制限酵素修飾酵素遺伝子の自己増殖の発見"海洋. 号外33号. 128-136 (2003)
[Publications] I.Kobayashi: "Addiction as a principle of symbiosis of genetic elements in the genome --- restriction enzymes, chromosomes and mitochondria."Endosymbiosis and Eukaryotic Organelles. M.Sugiura, R.Oelmueller, J.Obokata, Eds.(Logos Verlag Berlin, Berlin). (刊行中).
[Publications] I.Kobayashi: "Genetic addiction --- a principle in symbiosis of genes in a genome."Plasmid Biology. G.Phillips and B.Funnell, Eds.(ASM Press, Washington, D.C.). (刊行中).
[Publications] I.Kobayashi: "Restriction-modification systems as minimal forms of life."Restriction endonucleases. A Pingoud (ed.) (Springer-Verlag, Berlin). 19-62 (2003)
[Publications] I.Kobayashi: "Life cycle of restriction-modification systems, powers in genome evolution."Genome Science --- Towards a new paradigm? H.Yoshikawa, N.Ogasawara, N.Satoh, Eds.(Elsevier Science, Amsterdam). 199-202 (2003)
[Publications] 小林一三, 広瀬彩: "バクテリオファージ・ラムダ、細胞生物学事典"朝倉書店(刊行中).

2003 Fiscal Year Annual Research Report

利己的な制限酵素修飾酵素遺伝子のゲノム進化での役割に関する比較ゲノムと実験進化

Principal Investigator

小林 一三 東京大学, 医科学研究所, 助教授 (30126057)

Research Products

[Publications] A.Ino, S.Yamamoto, Y.Kaneda, I.Kobayashi: "Somatic gene targeting with RNA/DNA chimeric oligonucleotides : an analysis with a sensitive reporter mouse system."J.Gene Medicine. (刊行中).

[Publications] Y.Fujitani, I.Kobayashi: "Random-walkers active in genetic recombination."Biology International. (刊行中).

[Publications] K.Kusano, Y.Asami, A.Fujita, M.Tanokura, I.Kobayashi: "Type I restriction enzyme with RecA protein promotes illegitimate recombination."Plasmid. 50. 202-212 (2003)

[Publications] N.Handa, I.Kobayashi: "Accumulation of large non-circular forms of the chromosome in recombination-defective mutants of Escherichia coli."BMC Molecular Biology. 4. 5 (2003)

[Publications] R.J.Roberts, et al.: "A nomenclature for restriction enzymes, DNA methyltransferases, homing endonucleases and their genes."Nucleic Acids Res. 31. 1805-1812 (2003)

[Publications] M.Sadykov, Y.Asami, H.Niki, N.Handa, M.Itaya, M.Tanokura, I.Kobayashi: "Multiplication of a restriction modification gene complex."Mol.Microbiol. 48. 417-427 (2003)

[Publications] N.Takahashi, H.Yoshikura, I.Kobayashi: "An Escherichia coli strain, BJ5183, that shows highly efficient conservative (two-progeny) DNA double-strand break repair of restriction breaks."Gene. 303. 89-97 (2003)

[Publications] Y.Fujitani, I.Kobayashi: "Asymmetric Random Walk in a Reaction Intermediate of Homologous Recombination."J.Theor.Biol.. 220. 359-370 (2003)

[Publications] 小林一三, 石川健: "制限酵素修飾酵素遺伝子の自己増殖の発見"海洋. 号外33号. 128-136 (2003)

[Publications] I.Kobayashi: "Addiction as a principle of symbiosis of genetic elements in the genome --- restriction enzymes, chromosomes and mitochondria."Endosymbiosis and Eukaryotic Organelles. M.Sugiura, R.Oelmueller, J.Obokata, Eds.(Logos Verlag Berlin, Berlin). (刊行中).

[Publications] I.Kobayashi: "Genetic addiction --- a principle in symbiosis of genes in a genome."Plasmid Biology. G.Phillips and B.Funnell, Eds.(ASM Press, Washington, D.C.). (刊行中).

[Publications] I.Kobayashi: "Restriction-modification systems as minimal forms of life."Restriction endonucleases. A Pingoud (ed.) (Springer-Verlag, Berlin). 19-62 (2003)

[Publications] I.Kobayashi: "Life cycle of restriction-modification systems, powers in genome evolution."Genome Science --- Towards a new paradigm? H.Yoshikawa, N.Ogasawara, N.Satoh, Eds.(Elsevier Science, Amsterdam). 199-202 (2003)

[Publications] 小林一三, 広瀬彩: "バクテリオファージ・ラムダ、細胞生物学事典"朝倉書店(刊行中).

小林一三東京大学, 医科学研究所, 助教授 (30126057)