Novel plasmid marker rescue transformation system for molecular cloning in Bacillus subtilis enabling direct selection of recombinants

Molecular & General Genetics : MGG
P HaimaG Venema

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.

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Citations

Mar 8, 2012·Applied Microbiology and Biotechnology·Ljubica VojcicUlrich Schwaneberg
Sep 1, 1991·Research in Microbiology·S BronP Haima
Jan 1, 1994·Society for Applied Bacteriology Symposium Series·P J HillG S Stewart
Apr 27, 2010·Applied and Environmental Microbiology·Leonard H DamelinCaroline T Tiemessen
Jan 3, 2001·Molecular Microbiology·C A PritzlaffV Nizet
Apr 14, 1999·Molecular Microbiology·N N BaranovaA A Neyfakh

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