title: Novel plasmid marker rescue transformation system for molecular cloning in Bacillus subtilis enabling direct selection of recombinants.
abstract: A versatile plasmid marker rescue transformation system was developed for homology-facilitated cloning in Bacillus subtilis. It is based on the highly efficient host-vector system 6GM15-pHPS9, which allows the direct selection of recombinants by means of beta-galactosidase alpha-complementation. The system offers several advantages over previously described cloning systems: (1) the convenient direct selection of recombinants; (2) the ability to effectively transform B. subtilis competent cells with plasmid monomers, which allows the forced cloning of DNA fragments with high efficiency; (3) the availability of 6 unique target sites, which can be used for direct clone selection, SphI, NdeI, NheI, BamHI, SmaI and EcoRI; and (4) the rapid segregational loss of the helper plasmid from the transformed cells.
authors.affiliation: Department of Microbiology, Center of Biological Sciences, Haren Groningen, The Netherlands.
authors.forename: P
authors.initials: P
authors.lastname: Haima
authors.forename: S
authors.initials: S
authors.lastname: Bron
authors.forename: G
authors.initials: G
authors.lastname: Venema
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis; D004274:DNA, Recombinant / Q000032:analysis*; D005816:Genetic Complementation Test; D005819:Genetic Markers; D010957:Plasmids; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014169:Transformation, Bacterial
chemical_list: D004269:DNA, Bacterial; D004274:DNA, Recombinant; D005819:Genetic Markers