title: An optimized technique for rapid genome modifications of Bacillus subtilis.
abstract: The transformation efficiency of naturally competent Bacillus subtilis cells can be significantly increased using β recombinase binding sequences, as revealed by the results of this study. Plasmids containing different variations of these so called six-site-marker-cassettes were investigated. Furthermore, an optimized protocol for knock-out or knock-in mutations combining the Cre-lox-system and the six-sites is presented, which can be used for multiple genome modifications of B. subtilis.
authors.affiliation: Institute of Pharmacy, Dept. of Pharmaceutical Biotechnology, Ernst-Moritz-Arndt-University, 17487 Greifswald, Germany.
authors.forename: Jana
authors.initials: J
authors.lastname: Kumpfmüller
authors.forename: Johannes
authors.initials: J
authors.lastname: Kabisch
authors.forename: Thomas
authors.initials: T
authors.lastname: Schweder
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics*; D055879:Gene Knock-In Techniques / Q000379:methods; D055786:Gene Knockout Techniques / Q000379:methods; D005827:Genetics, Microbial / Q000379:methods*; D014169:Transformation, Bacterial
keywords: Bacillus subtilis; CAA; CFU; Gene library; Natural competence; OD; PCR; SSS; Six sites; TM; Transformation efficiency; Wild type strain; base pairs; bp; casamino acids; colony forming units; optical density; polymerase chain reaction; six-site-spectinomycin resistance gene; transformation medium