A new simple method to introduce marker-free deletions in the Bacillus subtilis genome.
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.
Morimoto
Takuya
T
Graduate School of Information Science, Nara Institute of Science and Technology, Nara 630-0101, Japan.
Ara
Katsutoshi
K
Ozaki
Katsuya
K
Ogasawara
Naotake
N
D001412:Bacillus subtilis / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics; D004722:Endoribonucleases / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics; D017353:Gene Deletion; D005819:Genetic Markers / Q000235:genetics; D016680:Genome, Bacterial / Q000235:genetics*; D011995:Recombination, Genetic
D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D005819:Genetic Markers; C422955:MazF protein, E coli; D004722:Endoribonucleases