A simple method for introducing marker-free deletions in the bacillus subtilis genome

Takuya Morimoto, Katsutoshi Ara, Katsuya Ozaki, Naotake Ogasawara

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Citations (Scopus)

Abstract

A genetic tool for introducing marker-free deletions is essential for multiple manipulations of genomes. We have developed a simple and efficient method for creating 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 counterselection tool.

Original languageEnglish
Title of host publicationStrain Engineering
Subtitle of host publicationMethods and Protocols
EditorsJames Williams
Pages345-358
Number of pages14
DOIs
Publication statusPublished - 2011 Sep 23
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume765
ISSN (Print)1064-3745

Fingerprint

Endoribonucleases
Bacillus subtilis
Genome
Escherichia coli
Polymerase Chain Reaction
Messenger RNA
Genes

Keywords

  • Bacillus subtilis
  • Marker-free mutant
  • MazF

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Morimoto, T., Ara, K., Ozaki, K., & Ogasawara, N. (2011). A simple method for introducing marker-free deletions in the bacillus subtilis genome. In J. Williams (Ed.), Strain Engineering: Methods and Protocols (pp. 345-358). (Methods in Molecular Biology; Vol. 765). https://doi.org/10.1007/978-1-61779-197-0_20

A simple method for introducing marker-free deletions in the bacillus subtilis genome. / Morimoto, Takuya; Ara, Katsutoshi; Ozaki, Katsuya; Ogasawara, Naotake.

Strain Engineering: Methods and Protocols. ed. / James Williams. 2011. p. 345-358 (Methods in Molecular Biology; Vol. 765).

Research output: Chapter in Book/Report/Conference proceedingChapter

Morimoto, T, Ara, K, Ozaki, K & Ogasawara, N 2011, A simple method for introducing marker-free deletions in the bacillus subtilis genome. in J Williams (ed.), Strain Engineering: Methods and Protocols. Methods in Molecular Biology, vol. 765, pp. 345-358. https://doi.org/10.1007/978-1-61779-197-0_20
Morimoto T, Ara K, Ozaki K, Ogasawara N. A simple method for introducing marker-free deletions in the bacillus subtilis genome. In Williams J, editor, Strain Engineering: Methods and Protocols. 2011. p. 345-358. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-61779-197-0_20
Morimoto, Takuya ; Ara, Katsutoshi ; Ozaki, Katsuya ; Ogasawara, Naotake. / A simple method for introducing marker-free deletions in the bacillus subtilis genome. Strain Engineering: Methods and Protocols. editor / James Williams. 2011. pp. 345-358 (Methods in Molecular Biology).
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