Development of a genetic transformation system for benzene-tolerant rhodococcus opacus strains
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概要
- 論文の詳細を見る
Rhodococcus opacus B-4 and B-9 are tolerant to various organic solvents including benzene, toluene, ethylbenzene, xylenes and styrene, and are suitable bacterial hosts for the production of chemical products from hydrophobic substrates. A 4.4-kb endogenous plasmid (pKNR01) was isolated from R. opacus B-4 and sequenced completely. Plasmid pKNR01 encodes proteins that share similarity to replication proteins from the enteric bacterial and actinomycete θ-replication plasmids. A 7.4-kb chimeric plasmid, designated pKNR01.1, was constructed by fusing XhoI-digested pKNR01 and Escherichia coli vector pSTV28. Plasmid pKNR01.1 had the ability to replicate in B-4 and B-9. A protocol for transformation of B-9 by electroporation was optimized employing pKNR01.1. Frequencies of 4.1×105 transformants per μg of plasmid DNA were obtained for B-9 cells, whereas B-4 harboring naturally occurring pKNR01 was transformed at lower frequencies (approximately 1×104 transformants per μg of plasmid DNA). Deletion analysis of pKNR01.1 showed that the 1.9-kb SphI-XhoI region containing the repA and repB genes and the 0.6-kb region upstream of repA was essential for plasmid maintenance in R. opacus strains.
- 社団法人日本生物工学会の論文
- 2005-04-25
著者
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Ohtake Hisao
Department of Biotechnology, Graduate School of Engineering, Osaka University
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Kato Junichi
Department Of Cardiology Asahikawa Medical College
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KURODA Akio
Department of Molecular Biotechnology, Hiroshima University
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Ohtake H
Department Of Biotechnology Osaka University
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Ohtake Hisao
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Ohtake H
Institute Of Applied Microbiology University Of Tokyo
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Ohtake Hisao
Institute Of Applied Microbiology The University Of Tokyo
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Ohtake Hisao
Department Of Fermentation Technology Hiroshima University
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Kato J
Department Of Molecular Biotechnology Hiroshima University
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IKEDA TSUKASA
Department of Fermentation Technology, Hiroshima University
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TAKIGUCHI NOBORU
Department of Fermentation Technology, Hiroshima University
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Okumura H
Electrotechnical Lab. Ibaraki Jpn
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NA KYUNG
Department of Molecular Biotechnology, Hiroshima University
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NAGAYASU KAN
Department of Molecular Biotechnology, Hiroshima University
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Ohtake Hisao
大阪大学 工学研究科生命先端工学専攻
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Na Kyung
Department Of Molecular Biotechnology Graduate School Of Advanced Sciences Of Matter Hiroshima Unive
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Kuroda A
Department Of Molecular Biotechnology Hiroshima University
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Kuroda Akio
Department Of Molecular Biotechnology Hiroshima University
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Nagayasu Kan
Department Of Molecular Biotechnology Hiroshima University
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Kato Junichi
Department Of Agricultural Biological Chemistry College Of Bioresource Sciences Nihon University
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Kato J
Department Of Chemical Engineering Faculty Of Engineering Shizuoka University
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Ikeda Tsukasa
Department Of Applied Chemistry Utsunomiya University
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Ikeda T
Dep. Of Material And Environmental Chemistry Graduate School Of Engineering Utsunomiya Univ.
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Takiguchi Noboru
Department Of Molecular Biotechnology Hiroshima University
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Takiguchi Noboru
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Kato Junichi
Department Of Agricultural Chemistry University Of Tokyo
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Ikeda T
Osaka Medical Coll. Osaka
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Kuroda Akio
Department Of Chemistry Faculty Of Science Nagoya University
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Na Kyung-Su
Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University
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Okumura Hajime
Department of Biotechnology, Graduate School of Engineering, Osaka University
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