Genes & Genetic Systems
Online ISSN : 1880-5779
Print ISSN : 1341-7568
ISSN-L : 1341-7568
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A new simple method to introduce marker-free deletions in the Bacillus subtilis genome
Takuya MorimotoKatsutoshi AraKatsuya OzakiNaotake Ogasawara
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2009 年 84 巻 4 号 p. 315-318

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A genetic tool to introduce marker-free deletions is essential for multiple manipulations of genomes. We report a simple and efficient method to create marker-free deletion mutants of Bacillus subtilis through transformation with recombinant PCR products, using the Escherichia coli mazF gene encoding an endoribonuclease that cleaves free mRNAs as a counter-selection tool. Our method will be applicable to any bacterium in which introduction of the mazF cassette into the genome by double crossover homologous recombination is possible.
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© 2009 by The Genetics Society of Japan
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