Stable Positional Cloning of Long Continuous DNA in the <i>Bacillus subtilis</i> Genome Vector
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- Itaya Mitsuhiro
- Mitsubishi Kagaku Institute of Life Sciences
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- Fujita Kyoko
- Mitsubishi Kagaku Institute of Life Sciences
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- Ikeuchi Masahiko
- Department of Life Sciences (Biology), University of Tokyo
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- Koizumi Maki
- Mitsubishi Kagaku Institute of Life Sciences
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- Tsuge Kenji
- Mitsubishi Kagaku Institute of Life Sciences
Bibliographic Information
- Other Title
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- Stable Positional Cloning of Long Continuous DNA in the Bacillus subtilis Genome Vector
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Abstract
Direct cloning of a long continuous genome segment in a Bacillus subtilis genome vector was demonstrated for the first time. Two small DNA fragments had to be installed in the vector prior to cloning. The DNA between these two fragments was cloned via homologous recombination. The efficiency of cloning was estimated using the 3, 573-kb genome of a cyanobacterium, Synechocystis sp. PCC 6803. Recombinants were selected using the internal selection system of the Bacillus genome vector or with the antibiotic resistance marker in the cyanobacterial genome. Designated genomic segments as large as 77-kb were cloned by means of a single procedure. Cloning efficiency is affected by the molecular weight of the donor DNA and the size of the DNA to be cloned. The method is suitable for direct target cloning of large-sized DNA.
Journal
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- The Journal of Biochemistry
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The Journal of Biochemistry 134 (4), 513-519, 2003
The Japanese Biochemical Society
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Details 詳細情報について
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- CRID
- 1390001204964427648
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- NII Article ID
- 130003534594
- 10012058225
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- NII Book ID
- AA00694073
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- COI
- 1:CAS:528:DC%2BD3sXhtVWgtbrN
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- ISSN
- 17562651
- 0021924X
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- NDL BIB ID
- 6732796
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- PubMed
- 14607977
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- Abstract License Flag
- Disallowed