A new simple method to introduce marker-free deletions in the Bacillus subtilis genome

Takuya Morimoto, Katsutoshi Ara, Katsuya Ozaki, Naotake Ogasawara

Research output: Contribution to journalArticle

29 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)315-318
Number of pages4
JournalGenes and Genetic Systems
Issue number4
Publication statusPublished - 2009 Dec 1
Externally publishedYes



  • Bacillus subtilis
  • Marker-free mutant
  • mazF

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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