A Simple Method for Multiple Modification of the<i>Escherichia coli</i>K-12 Chromosome
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- MIZOGUCHI Hiroshi
- Biofrontier Laboratories, Kyowa Hakko Kogyo Co., Ltd.
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- TANAKA-MASUDA Kimie
- Biofrontier Laboratories, Kyowa Hakko Kogyo Co., Ltd.
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- MORI Hideo
- Biofrontier Laboratories, Kyowa Hakko Kogyo Co., Ltd.
書誌事項
- タイトル別名
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- A Simple Method for Multiple Modification of the Escherichia coli K-12 Chromosome
- A simple method for multiple modification of the <italic>Escherichia coli</italic> K-12 chromosome
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説明
We developed a simple method of generating markerless deletions in the Escherichia coli chromosome. The method consists of two recombination events stimulated by λ Red recombinase. The first recombination replaced a target region with a marker cassette and the second then eliminated the marker cassette. The marker cassette included an antibiotic resistant gene and a negative selection marker (Bacillus subtilis sacB). Since sacB makes E. coli sensitive to sucrose, a markerless deletion strain was successfully selected using its sucrose-resistant phenotype. To stimulate these recombination events, 1-kbp homologous sequences adjacent to the target region were connected to both ends of the marker cassette or connected to each other by PCR. The average efficiency of the recombinations was 24% and 93% respectively. Eliminating the marker cassette with a fragment including an additional sequence, insertion was also possible. This markerless deletion method should be useful in creating a highly modified E. coli chromosome.
収録刊行物
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- Bioscience, Biotechnology, and Biochemistry
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Bioscience, Biotechnology, and Biochemistry 71 (12), 2905-2911, 2007
公益社団法人 日本農芸化学会
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詳細情報 詳細情報について
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- CRID
- 1390001206477451392
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- NII論文ID
- 10027521760
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- NII書誌ID
- AA10824164
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- ISSN
- 13476947
- 09168451
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- NDL書誌ID
- 9326458
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
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- 抄録ライセンスフラグ
- 使用不可