Abstract
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 language | English |
|---|---|
| Pages (from-to) | 315-318 |
| Number of pages | 4 |
| Journal | Genes and Genetic Systems |
| Volume | 84 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - 2009 Dec 1 |
| Externally published | Yes |
Keywords
- Bacillus subtilis
- Marker-free mutant
- mazF
ASJC Scopus subject areas
- Molecular Biology
- Genetics
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Morimoto, T., Ara, K., Ozaki, K., & Ogasawara, N. (2009). A new simple method to introduce marker-free deletions in the Bacillus subtilis genome. Genes and Genetic Systems, 84(4), 315-318. https://doi.org/10.1266/ggs.84.315