title: A simple method for introducing marker-free deletions in the Bacillus subtilis genome.
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.
authors.affiliation: Graduate School of Information Science, Nara Institute of Science and Technology, Takayama, Ikoma, Nara, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Morimoto
authors.forename: Katsutoshi
authors.initials: K
authors.lastname: Ara
authors.forename: Katsuya
authors.initials: K
authors.lastname: Ozaki
authors.forename: Naotake
authors.initials: N
authors.lastname: Ogasawara
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics*; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D017353:Gene Deletion; D005819:Genetic Markers / Q000235:genetics; D016680:Genome, Bacterial / Q000235:genetics*; D016133:Polymerase Chain Reaction; D014169:Transformation, Bacterial / Q000235:genetics*
chemical_list: D004274:DNA, Recombinant; D005819:Genetic Markers; D004722:Endoribonucleases